Novel sulfonamide inhibitors of aspartyl protease

ABSTRACT

The present invention relates to a novel class of sulfonamides which are aspartyl protease inhibitors. In one embodiment, this invention relates to a novel class of HIV aspartyl protease inhibitors characterized by specific structural and physicochemical features. This invention also relates to pharmaceutical compositions comprising these compounds. The compounds and pharmaceutical compositions of this invention are particularly well suited for inhibiting HIV-1 and HIV-2 protease activity and consequently, may be advantageously used as anti-viral agents against the HIV-1 and HIV-2 viruses. This invention also relates to methods for inhibiting the activity of HIV aspartyl protease using the compounds of this invention and methods for screening compounds for anti-HIV activity.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to a novel class of sulfonamides which areaspartyl protease inhibitors. In one embodiment, this invention relatesto a novel class of HIV aspartyl protease inhibitors characterized byspecific structural and physicochemical features. This invention alsorelates to pharmaceutical compositions comprising these compounds. Thecompounds and pharmaceutical compositions of this invention areparticularly well suited for inhibiting HIV-1 and HIV-2 proteaseactivity and consequently, may be advantageously used as anti-viralagents against the HIV-1 and HIV-2 viruses. This invention also relatesto methods for inhibiting the activity of HIV aspartyl protease usingthe compounds of this invention and methods for screening compounds foranti-HIV activity.

BACKGROUND OF THE INVENTION

The human immunodeficiency virus (“HIV”) is the causative agent foracquired immunodeficiency syndrome (“AIDS”)—a disease characterized bythe destruction of the immune system, particularly of CD4⁺ T-cells, withattendant susceptibility to opportunistic infections—and its precursorAIDS-related complex (“ARC”)—a syndrome characterized by symptoms suchas persistent generalized lymphadenopathy, fever and weight loss.

As in the case of several other retroviruses, HIV encodes the productionof a protease which carries out post-translational cleavage of precursorpolypeptides in a process necessary for the formation of infectiousvirions (S. Crawford et al., “A Deletion Mutation in the 5′ Part of thepol Gene of Moloney Murine Leukemia Virus Blocks Proteolytic Processingof the gag and pol Polyproteins”, J. Virol., 53, p. 899 (1985)). Thesegene products include pol, which encodes the virion RNA-dependent DNApolymerase (reverse transcriptase), an endonuclease, HIV protease, andgag, which encodes the core-proteins of the virion (H. Toh et al.,“Close Structural Resemblance Between Putative Polymerase of aDrosophila Transposable. Genetic Element 17.6 and pol gene product ofMoloney Murine Leukemia Virus”, EMBO J., 4, p. 1267 (1985); L. H. Pearlet al., “A Structural Model for the Retroviral Proteases”, Nature, pp.329-351 (1987); M. D. Power et al., “Nucleotide Sequence of SRV-1, aType D Simian Acquired Immune Deficiency Syndrome Retrovirus”, Science,231, p. 1567 (1986)).

A number of synthetic anti-viral agents have been designed to targetvarious stages in the replication cycle of HIV. These agents includecompounds which block viral binding to CD4⁺ T-lymphocytes (for example,soluble CD4), and compounds which interfere with viral replication byinhibiting viral reverse transcriptase (for example, didanosine andzidovudine (AZT)) and inhibit integration of viral DNA into cellular DNA(M. S. Hirsh and R. T. D′Aqulia, “Therapy for Human ImmunodeficiencyVirus Infection”, N. Eng. J. Med., 328, p. 1686 (1993)). However, suchagents, which are directed primarily to early stages of viralreplication, do not prevent the production of infectious virions inchronically infected cells. Furthermore, administration of some of theseagents in effective amounts has led to cell-toxicity and unwanted sideeffects, such as anemia and bone marrow suppression.

More recently, the focus of anti-viral drug design has been to createcompounds which inhibit the formation of infectious virions byinterfering with the processing of viral polyprotein precursors.Processing of these precursor proteins requires the action ofvirus-encoded proteases which are essential for replication (Kohl, N. E.et al. “Active HIV Protease is Required for Viral Infectivity” Proc.Natl. Acad. Sci. USA, 85, p. 4686 (1988)). The anti-viral potential ofHIV protease inhibition has been demonstrated using peptidal inhibitors.Such peptidal compounds, however, are typically large and complexmolecules that tend to exhibit poor bioavailability and are notgenerally consistent with oral administration. Accordingly, the needstill exists for compounds that can effectively inhibit the action ofviral proteases, for use as agents for preventing and treating chronicand acute viral infections.

SUMMARY OF THE INVENTION

The present invention provides a novel class of compounds, andpharmaceutically acceptable derivatives thereof, that are useful asinhibitors of aspartyl proteases, in particular, HIV aspartyl protease.These compounds can be used alone or in combination with othertherapeutic or prophylactic agents, such as anti-virals, antibiotics,immunomodulators or vaccines, for the treatment or prophylaxis of viralinfection.

According to a preferred embodiment, the compounds of this invention arecapable of inhibiting HIV viral replication in human CD₄ ⁺ T-cells.These compounds are useful as therapeutic and prophylactic agents totreat or prevent infection by HIV-1 and related viruses which may resultin asymptomatic infection, AIDS-related complex (“ARC”), acquiredimmunodeficiency syndrome (“AIDS”), or similar disease of the immunesystem.

It is a principal object of this invention to provide a novel class ofsulfonamides which are aspartyl protease inhibitors, and particularly,HIV aspartyl protease inhibitors. This novel class of sulfonamides isrepresented by formula I:

wherein:

A is selected from the group consisting of H; Het; —R¹—Het; —R¹—C₁-C₆alkyl, which may be optionally substituted with one or more groupsselected from the group consisting of hydroxy, C₁-C₄ alkoxy, Het,—O-Het, —NR²—CO—N(R²)(R²) and —CO—N(R²)(R²); and —R¹—C₂-C₆ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het, —O-Het,—NR²—CO—N(R²)(R²) and —CO—N(R²)(R²);

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)— and—NR²—C(O)—C(O)—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, N(R²), O, S and S(O)_(n), wherein said heterocycle mayoptionally be benzofused; and wherein any member of said Het may beoptionally substituted with one or more substituents selected from thegroup consisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)(R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R², —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃, —NO₂, Ar and —O—Ar;

each R² is independently selected from the group consisting of H andC₁-C₃ alkyl optionally substituted with Ar;

B, when present, is —N(R²)—C(R³)(R³)—C(O)—;

x is 0 or 1;

each R³ is independently selected from the group consisting of H, Het,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl,wherein any member of said R³, except H, may be optionally substitutedwith one or more substituents selected from the group consisting of—OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR², —CO₂R²,NR²—C(O)—R²;

each n is independently 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with one or more groupsselected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₂-C₄ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar;C₃-C₆ cycloalkyl, which may be optionally substituted with or fused withAr; and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with Ar;

each Ar is independently selected from the group consisting of phenyl;3-6 membered carbocyclic ring and 5-6 membered heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) andN(R²), wherein said carbocyclic or heterocyclic ring may be saturated orunsaturated and optionally substituted with one or more groups selectedfrom the group consisting of oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)(R²), halo and —CF₃;

E is selected from the group consisting of Het; O-Het; Het-Het; —O—R³;—NR²R³; C₁-C₆ alkyl, which may be optionally substituted with one ormore groups selected from the group consisting of R⁴ and Het; C₂-C₆alkenyl, which may be optionally substituted with one or more groupsselected from the group consisting of R⁴ and Het; C₃-C₆ saturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and C₅-C₆ unsaturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², —S(O)₂—NHR², halo, —NR²—C(O)—R² and —CN.

It is a also an object of this invention to provide pharmaceuticalcompositions comprising the sulfonamides of formula I and methods fortheir use as inhibitors of HIV aspartyl protease.

It is a further object of this invention to provide a novel class of HIVaspartyl protease inhibitor compounds characterized by the followingnovel combination of structural and physicochemical features:

(1) a first and a second hydrogen bond acceptor moiety, at least one ofwhich is more highly polarizable than a carbonyl, said moieties beingthe same or different, and being capable of hydrogen bonding with thehydrogen atoms of the flap water molecule of an HIV aspartyl proteasewhen the compound is bound thereto;

(2) substantially hydrophobic moieties which associate with the P₁ andP₁′ binding pockets of said HIV aspartyl protease when the compound isbound thereto;

(3) a third hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease when the compound is boundthereto;

(4) an additional occupied volume of space of at least 100 Å³ when thecompound is bound to the active site of said HIV aspartyl protease, saidspace overlapping with the volume of space that would be filled by anative substrate of said HIV aspartyl protease or a nonhyrolyzableisostere thereof;

(5) a deformation energy of binding of the compound to said HIV aspartylprotease of not greater than 10 kcal/mole; and

(6) a neutral or favorable enthalpic contribution from the sum of allelectrostatic interactions between the compound and the protease whenthe compound is bound to said HIV aspartyl protease.

It is also an object of this invention to provide pharmaceuticalcompositions comprising compounds having the above-mentioned featuresand methods for their use as inhibitors of HIV aspartyl protease.

It is a further object of this invention to provide a method foridentification, design, or prediction of HIV aspartyl proteaseinhibitors comprising the steps of:

-   -   (a) selecting a candidate compound of defined chemical structure        containing a first and a second hydrogen bond acceptor moiety,        at least one of which is more highly polarizable than a        carbonyl, said moieties being the same or different; a third        hydrogen bonding moiety, which may be either a hydrogen bond        donor or acceptor; and at least two substantially hydrophobic        moieties;    -   (b) determining a low-energy conformation for binding of said        compound to the active site of an HIV aspartyl protease;    -   (c) evaluating the capability of said first and second hydrogen        bond acceptor moieties to form hydrogen bonds to the flap water        molecule of said HIV aspartyl protease when said compound is        bound thereto in said conformation;    -   (d) evaluating the capability of said substantially hydrophobic        moieties to associate with the P₁ and P₁′ binding pockets of        said HIV aspartyl protease when said compound is bound thereto        in said conformation;    -   (e) evaluating the capability of said third hydrogen bonding        moiety to form hydrogen bonds to Asp25 and Asp25′ of said HIV        aspartyl protease when said compound is bound thereto in said        conformation;    -   (f) evaluating the overlap of the occupied volume of said        compound when said compound is bound to said HIV aspartyl        protease in said conformation and the occupied volume of a        native substrate of HIV aspartyl protease or a nonhydrolyzable        isostere thereof, when said polypeptide is bound to said HIV        aspartyl protease;    -   (g) evaluating the deformation energy of binding of said        compound to said HIV aspartyl protease;    -   (h) evaluating the enthalpic contribution of the sum of all        electrostatic interactions between said compound and said HIV        aspartyl protease when said compound is bound thereto in said        conformation; and    -   (i) accepting or rejecting said candidate compound as an HIV        protease inhibitor based upon the determinations and evaluations        carried out in steps (b) through (h).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a stereo drawing of a low-energy conformation of Compound140, as predicted by computer-modelling.

FIG. 2 depicts a stereo drawing of the actual crystal structure ofCompound 140, as observed by X-ray crystallography.

FIG. 3 depicts a stereo drawing of the correlation between the predicted(thin line) and observed (thick line) conformation of Compound 140.

DETAILED DESCRIPTION OF THE INVENTION

In order that the invention herein described may be more fullyunderstood, the following detailed description is set forth. In thedescription, the following abbreviations are used:

Designation Reagent or Fragment Ac acetyl Me methyl Et ethyl Bzl benzylTrityl triphenylmethyl Asn D- or L-asparagine Ile D- or L-isoleucine PheD- or L-phenylalanine Val D- or L-valine Boc tert-butoxycarbonyl Cbzbenzyloxycarbonyl (carbobenzyloxy) Fmoc 9-fluorenylmethoxycarbonyl DCCdicyclohexylcarbodiimide DIC diisopropylcarbodiimide EDC1-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride HOBt1-hydroxybenzotriazole HOSu 1-hydroxysuccinimide TFA trifluoroaceticacid DIEA diisopropylethylamine DBU 1,8-diazabicyclo(5.4.0)undec-7-eneEtOAc ethyl acetate

The following terms are employed herein:

Unless expressly stated to the contrary, the terms “—SO₂—” and “—S(O)₂—”as used herein refer to a sulfone or sulfone derivative (i.e., bothappended groups linked to the S), and not a sulfinate ester.

For the compounds of formula I, and intermediates thereof, thestereochemistry of the explicitly shown hydroxyl is defined relative toD on the adjacent carbon atom, when the molecule is drawn in an extendedzig-zag representation (such as that drawn for compounds of formula XI,XV, XXII, XXIII and XXXI). If both OH and D reside on the same side ofthe plane defined by the extended backbone of the compound, thestereochemistry of the hydroxyl will be referred to as “syn”. If OH andD reside on opposite sides of that plane, the stereochemistry of thehydroxyl will be referred to as “anti”.

The term “heterocyclic” refers to a stable 5-7 membered monocycle or8-11 membered bicyclic heterocycle which is either saturated orunsaturated, and which may be optionally benzofused if monocyclic. Eachheterocycle consists of carbon atoms and from one to four heteroatomsselected from the group consisting of nitrogen, oxygen and sulfur. Asused herein, the terms “nitrogen and sulfur heteroatoms” include anyoxidized form of nitrogen and sulfur, and the quaternized form of anybasic nitrogen. The heterocyclic ring may be attached by any heteroatomof the cycle which results in the creation of a stable structure.Preferred heterocycles defined above include, for example,benzimidazolyl, imidazolyl, imidazolinoyl, imidazolidinyl, quinolyl,isoquinolyl, indolyl, pyridyl, pyrrolyl, pyrrolinyl, pyrazolyl,pyrazinyl, quinoxolyl, piperidinyl, morpholinyl, thiamorpholinyl, furyl,thienyl, triazolyl, thiazolyl, β-carbolinyl, tetrazolyl, thiazolidinyl,benzofuanoyl, thiamorpholinyl sulfone, benzoxazolyl, oxopiperidinyl,oxopyrroldinyl, oxoazepinyl, azepinyl, isoxazolyl, tetrahydropyranyl,tetrahydrofuranyl, thiadiazoyl, benzodioxolyl, thiophenyl,tetrahydrothiophenyl and sulfolanyl.

The terms “HIV protease” and “HIV aspartyl protease” are usedinterchangeably and refer to the aspartyl protease encoded by the humanimmunodeficiency virus type 1 or 2. In a preferred embodiment of thisinvention, these terms refer to the human immunodeficiency virus type 1aspartyl protease.

The term “hydrophobic” refers to a moiety which tends not to dissolvereadily in water and is often fat-soluble. Hydrophobic moieties include,but are not limited to, hydrocarbons, such as alkanes, alkenes, alkynes,cycloalkanes, cycloalkenes, cycloalkynes and aromatic hydrocarbons, suchas aryls, certain saturated and unsaturated heterocycles and moietiesthat are substantially similar to the side chains of hydrophobic naturaland unnatural α-amino acids, including valine, leucine, isoleucine,methionine, phenylalanine, α-amino isobutyric acid, alloisoleucine,tyrosine, and tryptophan.

The term “substantially hydrophobic” refers to a hydrophobic moietywhich may optionally contain polar atoms or groups in the region of themoiety which are solvent exposed when the compound is bound in theactive site of an aspartyl protease.

The term “linker moiety” refers to a group within a compound, said groupconsisting of a backbone of 1-6 atoms selected from the group consistingof C, N, O, S and P, said backbone being substituted with, fused to orotherwise associated with a substantially hydrophobic group capable ofassociating with the P₁ or P₁′ binding pocket of an HIV aspartylprotease when said compound is bound thereto. In alternative embodimentsof this invention, such linker moieties may optionally be substitutedwith a group or groups which occupy a volume of space overlapping withthe volume of space that would be filled by a native substrate of HIVaspartyl protease or a nonhydrolyzable isostere thereof.

The term “more highly polarizable than a carbonyl” refers to a moietyhaving a polarizability (α) greater than that of a carbonyl group of acorresponding aldehyde, ketone, ester or amide moiety.

The term “pharmaceutically effective amount” refers to an amounteffective in treating HIV infection in a patient. The term“prophylactically effective amount” refers to an amount effective inpreventing HIV infection in a patient. As used herein, the term“patient” refers to a mammal, including a human.

The term “pharmaceutically acceptable carrier or adjuvant” refers to anon-toxic carrier or adjuvant that may be administered to a patient,together with a compound of this invention, and which does not destroythe pharmacological activity thereof.

As used herein, the compounds of this invention, including the compoundsof formula I, are defined to include pharmaceutically acceptablederivatives thereof. A “pharmaceutically acceptable derivative” meansany pharmaceutically acceptable salt, ester, or salt of such ester, of acompound of this invention or any other compound which, uponadministration to a recipient, is capable of providing (directly orindirectly) a compound of this invention or an anti-virally activemetabolite or residue thereof.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acids includehydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric,maleic, phosphoric, glycollic, lactic, salicylic, succinic,toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic,benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids.Other acids, such as oxalic, while not in themselves pharmaceuticallyacceptable, may be employed in the preparation of salts useful asintermediates in obtaining the compounds of the invention and theirpharmaceutically acceptable acid addition salts.

Salts derived from appropriate bases include alkali metal (e.g.,sodium), alkaline earth metal (e.g., magnesium), ammonium and N—(C₁₋₄alkyl)₄ ⁺ salts.

The term “thiocarbamates” refers to compounds containing the functionalgroup N—SO₂—O.

The compounds of this invention contain one or more asymmetric carbonatoms and thus occur as racemates and racemic mixtures, singleenantiomers, diastereomeric mixtures and individual diastereomers. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. Each stereogenic carbon may be of the R or Sconfiguration. The explicitly shown hydroxyl is also preferred to be synto D, in the extended zig-zag conformation between the nitrogens shownin compounds of formula I.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and administration to a mammalby methods known in the art. Typically, such compounds are stable at atemperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

The compounds of the present invention may be used in the form of saltsderived from inorganic or organic acids. Included among such acid salts,for example, are the following: acetate, adipate, alginate, aspartate,benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate,camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate,ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate,pectinate, persulfate, 3-phenylpropionate, picrate, pivalate,propionate, succinate, tartrate, thiocyanate, tosylate and undecanoate.

This invention also envisions the quaternization of any basicnitrogen-containing groups of the compounds disclosed herein. The basicnitrogen can be quaternized with any agents known to those of ordinaryskill in the art including, for example, lower alkyl halides, such asmethyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkylsulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; longchain halides such as decyl, lauryl, myristyl and stearyl chlorides,bromides and iodides; and aralkyl halides including benzyl and phenethylbromides. Water or oil-soluble or dispersible products may be obtainedby such quaternization.

The novel sulfonamides of this invention are those of formula I:

wherein:

A is selected from the group consisting of H; Het; —R¹-Het; —R¹—C₁-C₆alkyl, which may be optionally substituted with one or more groupsselected from the group consisting of hydroxy, C₁-C₄ alkoxy, Het,—O-Het, —NR²—CO—N(R²)(R²) and —CO—N(R²)(R²); and —R¹—C₂-C₆ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het, —O-Het,—NR²—CO—N(R²)(R²) and —CO—N(R²)(R²);

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—C(O)—, —O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)— and—NR²—C(O)—C(O)—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, N(R²), O, S and S(O)_(n), wherein said heterocycle mayoptionally be benzofused; and wherein any member of said Het may beoptionally substituted with one or more substituents selected from thegroup consisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)(R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R², —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃₁—NO₂, Ar and —O—Ar;

each R² is independently selected from the group consisting of H andC₁-C₃ alkyl optionally substituted with Ar;

B, when present, is —N(R²)—C(R³)(R³)—C(O)—;

x is 0 or 1;

each R³ is independently selected from the group consisting of H, Het,C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl,wherein any member of said R³, except H, may be optionally substitutedwith one or more substituents selected from the group consisting of—OR², —C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR², —CO₂R²,NR²—C(O)—R²;

each n is independently 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with one or more groupsselected from C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₂-C₄ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar;C₃-C₆ cycloalkyl, which may be optionally substituted with or fused withAr; and C₅-C₆ cycloalkenyl, which may be optionally substituted with orfused with Ar;

each Ar is independently selected from the group consisting of phenyl;3-6 membered carbocyclic ring and 5-6 membered heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) andN(R²), wherein said carbocyclic or heterocyclic ring may be saturated orunsaturated and optionally substituted with one or more groups selectedfrom the group consisting of oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)(R²), halo and —CF₃;

E is selected from the group consisting of Het; O-Het; Het-Het; —O—R³;—NR²R³; C₁-C₆ alkyl, which may be optionally substituted with one ormore groups selected from the group consisting of R⁴ and Het; C₂-C₆alkenyl, which may be optionally substituted with one or more groupsselected from the group consisting of R⁴ and Het; C₃-C₆ saturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and C₅-C₆ unsaturatedcarbocycle, which may optionally be substituted with one or more groupsselected from the group consisting of R⁴ and Het; and

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², S(O)₂—NHR², halo, —NR²—C(O)—R² and —CN.

Except where expressly provided to the contrary, as used herein, thedefinitions of variables A, R¹-R⁴, Het, B, x, n, D, D′, Ar and E are tobe taken as they are defined above for the compounds of formula I.

According to one embodiment of this invention, a subclass of compoundsare those compounds of formula I, and pharmaceutically acceptable saltsthereof, wherein:

A is selected from the group consisting of H; —R¹—Het; —R¹—C₁-C₆ alkyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het and —O-Het; and—R¹—C₂-C₆ alkenyl, which may be optionally substituted with one or moregroups selected from hydroxy, C₁-C₄ alkoxy, Het and —O-Het;

each R¹ is independently selected from the group consisting of —C(O)—,—S(O)₂—, —C(O)—C(O)—, —O—CO—, —O—S(O)₂— and —NR²—;

each Het is independently selected from the group consisting of C₃-C₇cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturatedor unsaturated heterocycle, containing one or more heteroatoms selectedfrom N, O and S, which may optionally be benzofused; wherein any memberof said Het may be optionally substituted with one or more substituentsselected from the group consisting of oxo, —OR², —R², —N(R²)₂″—R²—OH,—CN, —CO₂R², —C(O)—N(R²)₂ and —S(O)₂—N(R²)₂;

each R² is independently selected from the group consisting of H andC₁-C₃ alkyl;

B, when present, is —NH—CH(R³)—C(O)—;

x is 0 or 1;

R³ is selected from the group consisting of Het, C₁-C₆ alkyl, C₂-C₆alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl, wherein any member ofsaid R³ may be optionally substituted with one or more substituentsselected from the group consisting of —OR², —C(O)—NH—R²,—S(O)_(n)—N(R²)₂, Het and —CN;

n is 1 or 2;

D and D′ are independently selected from the group consisting of Ar;C₁-C₄ alkyl, which may be optionally substituted with C₃-C₆ cycloalkylor Ar; C₂-C₄ alkenyl, which may be optionally substituted with C₃-C₆cycloalkyl or Ar; C₃-C₆ cycloalkyl, which may be optionally substitutedor fused with Ar; and C₅-C₆ cycloalkenyl, which may be optionallysubstituted or fused with Ar; with the proviso that when D is attachedto N, D may not be methyl or C₂ alkenyl;

Ar is selected from the group consisting of phenyl; 3-6 memberedcarbocyclic ring and 5-6 membered heterocyclic ring containing one ormore heteroatoms selected from O, N and S, wherein said carbocyclic orheterocyclic ring may be saturated or unsaturated and optionallysubstituted with one or more groups selected from the group consistingof oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)R², —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)₂, halo and —CF₃;

E is selected from the group consisting of Het; —O—R³; —NR²R⁵; C₁-C₆alkyl, which may be optionally substituted with one or more R⁴ or Het;C₂-C₆ alkenyl, which may be optionally substituted with one or more R⁴or Het; C₃-C₆ saturated carbocycle, which may optionally be substitutedwith one or more R⁴ or Het; and C₅-C₆ unsaturated carbocycle, which mayoptionally be substituted with one or more R⁴ or Het;

each R⁴ is independently selected from the group consisting of —OR²,—C(O)—NHR², —S(O)₂—NHR², halo and —CN; and

each R⁵ is independently selected from the group consisting of H and R³,with the proviso that at least one R⁵ is not H.

A preferred subclass of compounds of this invention are those compoundsof formula I having a molecular weight of less than about 700 g/mole.More preferably, the subclass of compounds of formula I have a molecularweight of less than about 600 g/mole.

Other preferred subclasses of this invention are those compounds offormulas XXII, XXIII and XXXI:

wherein A, R³, Het, D, D′, x and E are as defined above for compounds offormula I. For ease of reference, the two R³ moieties present in formulaXXXI have been labeled R³ and R^(3′).

For compounds of formula XXII, most preferred compounds are thosewherein A is R¹-Het and D′ is C₁-C₃ alkyl or C₃ alkenyl, wherein saidalkyl or alkenyl may optionally be substituted with one or more groupsselected from the group consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar andAr (with all other variables being defined as above for compounds offormula I). For compounds of formula XXIII, most preferred compounds arethose wherein R³ is C₁-C₆ alkyl, C₂-C₆ alkenyl, C₅-C₆ cycloalkyl, C₅-C₆cycloalkenyl or a 5-6 membered saturated or unsaturated heterocycle,wherein any member of said R³ may be optionally substituted with one ormore substituents selected from the group consisting of —OR²,—C(O)—NH—R², —S(O)_(n)N(R²)(R²), Het, —CN, —SR², —C(O)₂R² andNR²—C(O)—R² and D′ is C₁-C₃ alkyl or C₃ alkenyl, wherein said alkyl oralkenyl may optionally be substituted with one or more groups selectedfrom the group consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar and Ar (withall other variables being defined as above for compounds of formula I).

For compounds of formula XXXI, most preferred compounds are thosewherein A is R¹-Het, each R³ is independently C₁-C₆ alkyl which may beoptionally substituted with a substituent selected from the groupconsisting of —OR², —C(O)—NH—R², —S(O)_(n)N(R²)(R²), Het, —CN, —SR²,—CO₂R² and —NR²—C(O)—R²; D′ is C₁-C₄ alkyl, which may be optionallysubstituted with a group selected from the group consisting of C₃-C₆cycloalkyl, —OR², —O—Ar; and E is Het, Het-Het and —NR²R³.

Sulfonamides of this invention include the following specific compoundscontained in Tables I-VI. In Tables I-IV and VI, A is attached throughthe right-most bond, unless otherwise expressly noted. All othersubstituents in Tables I-VI are attached via the left-most bond, unlessotherwise expressly noted.

TABLE I

COMPOUND A R³ D′ E 1

2

3

4

5

6

7

8

CH₃ 9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

TABLE II

COMPOUND A D′ E 27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

CH₃

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

CH₃

127

CH₃

128

CH₃

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

TABLE III

COMPOUND A D′ E 186

187

188

TABLE IV

COMPOUND A 189

190

TABLE V

COMPOUND E 191

192

193

194

TABLE VI

COMPOUND D D′ 195

196

Preferred compounds of this invention are*: * As can be appreciated bythose of ordinary skill in the art, many different conventions are usedin naming chemical compounds. Because of possible discrepencies in theart of chemical nomenclature, the structures shown in Tables I-VI hereinare controlling for the definition of compounds 1-195 and 1001-1015 ofthis invention.

-   (S)-N-1-(3-((3-Acetylamino-4-fluoro-benzenesulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    and    (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compounds 2);-   (S)-N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 5);-   (S)-N-1-(1-Benzyl-3-(benzyl-(5-isoxazol-3-yl-thiophene-2-sulfonyl)-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 6);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-benzyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 9);-   N-1-(1-(S)-Benzyl-3-(benzyl-(3-sulfamoyl-benzenesulfonyl)-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 10);-   (S)-N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 12);-   (S)-N-1-(3-((4-Benzenesulfonyl-thiophene-2-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 13);-   (S)-N-1-(1-(S)-Benzyl-3-((4-fluoro-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 14);-   (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 15);-   (S)-N-1-(3-((3-Acetylamino-4-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 16);-   (S)-N-1-(1-(S)-Benzyl-3-((4-acetylamino-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 17);-   (S)-N-1-(3-((5-Acetylamino-3-methyl-thiophene-2-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 18);-   (S)-N-1-(3-((3-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 19);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 20);-   N-1-((1S-2    syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfonylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 21);-   N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinamide    (compound 22);-   N-1-(3-((4-Acetylamino-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-4-yl-methoxycarbonyl)-succinamide    (compound 23);-   N-1-(3-((4-Fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-(pyridin-2-yl-methoxycarbonyl)-succinamide    (compound 26);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 35);-   3,4-Dichloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 37);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 44);-   2,4-Dimethyl-thiazole-5-sulfonic    acid-(1,1-dimethyl-ethoxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-amide (compound 46);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 48);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    and 4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compounds 52);-   Benzo(1,2,5)oxadiazole-5-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-isobutylamide    (compound 66);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl-phenyl)-acetamide    and N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compounds 86);-   N-(2-Fluoro-5-(((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 88);-   N-(3-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 91);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((R)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 93);-   N-(4-(((syn)-2-Hydroxy-(S)-4-phenyl-3-((tetrahydro-furan-(R)-3-yl)-oxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 94);-   4-Fluoro-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(R)-3-ylmethoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    and 4-Fluoro-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((tetrahydro-furan-(S)-3-ylmethoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compounds 97);-   4-Fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 98);-   4-Chloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-benzenesulfonamide    (compound 99);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 100);-   4-Fluoro-N-(2-(syn)-hydroxy-3-((2-oxazolidon-(S)-4-yl)-methoxycarbonylamino)-4-(S)-phenyl-butyl)-N-isobutyl-benzenesulfonamide    (compound 109);-   Benzene-1,3-disulfonic acid 1-amide 3-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-(S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 112);-   Furan-3-sulfonic acid (2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 113);-   N-((3-Allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-N-cyclopentylmethyl-4-fluoro-benzenesulfonamide    (compound 114);-   N-Cyclopentylmethyl-N-((3-ethoxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-4-fluoro-benzenesulfonamide    (compound 115);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 116);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonyl)-butyl)-benzenesulfonamide    (compound 118);-   N-(4-(Cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide    (compound 125);-   3-Chloro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 138);-   4-Chloro-N-cyclopentylmethyl-N-(2-(syn)-hydroxy-3-((2-oxazolidon-4-(S)-yl-methyl)-oxycarbonylamino)-4-phenyl-butyl)-benzenesulfonamide    (compound 139);-   N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 140);-   N-((3-allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 141);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide    (compound 142);-   Pyridine-3-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide,    trifluoroacetic acid salt (compound 144);-   5-Isoxazol-3-yl-thiophene-2-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 145);-   N-(4-((3-(Allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-cyclopentylmethylsulfamoyl)-phenyl)-acetamide    (compound 146);-   N-(4-(Cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-sulfamoyl)-phenyl)-acetamide    (compound 147);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 148);-   Pyridine-3-sulfonic acid cyclopentylmethyl-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-amide    (compound 149);-   Piperidine-1-sulfonic acid ((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-amide    (compound 150);-   N-4-((2-(syn)-Hydroxy-3-((2-methoxymethyl-allyloxycarbonylamino)-4-(S)-phenyl-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 155);-   1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid    ((allyloxycarbonylamino)-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-cyclopentylmethyl-amide (compound    156);-   1-Acetyl-2,3-dihydro-1H-indole-6-sulfonic acid cyclopentylmethyl-((2    syn,    3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-amide    (compound 157);-   N-Cyclohexylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 158);-   N-Cyclohexylmethyl-4-fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 159);-   N-(4-(Cyclohexylmethyl)-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide    (compound 160);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-(pyridin-4-yl-methoxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 163);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((syn)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methyl-benzenesulfonamide    (compound 165);-   N-cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide    (compound 166);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-nitro-benzenesulfonamide    (compound 167);-   4-Amino-N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 168);-   N-Cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 169);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-nitro-benzensulfonamide    (compound 170);-   4-Amino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 171);-   2,4-Diamino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 173);-   4-Hydroxy-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 175);-   N-Cyclopentylmethyl-4-fluoro-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 182);-   3,4-Dichloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 183);-   Benzyloxycarbonyl-(L)-isoleucine-N-(5-((3-amino-(2    syn,3S)-2-hydroxy-4-phenyl-butyl)-isobutyl-sulfamoyl)-2-fluoro-phenyl)-acetamide    (compound 187);-   N-((2    syn,3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 195);

and compounds 1001 through 1015.

-   More preferred compounds of this invention are:-   (S)-N-1-(1-(S)-Benzyl-2-(syn)-hydroxyl-3-(isobutyl-(5-pyridin-2-yl-thiophene-2-sulfonyl)-amino)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 12);-   (S)-N-1-(1-(S)-Benzyl-3-((4-fluoro-benzenesulfonyl)-isobutyl-amino)-2-(syn)-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 14);-   (S)-N-1-(3-((4-Acetylamino-3-fluoro-benzenesulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 15);-   (S)-N-1-(3-((Benzo(1,2,5)oxadiazole-4-sulfonyl)-isobutyl-amino)-(1S,2    syn)-1-benzyl-2-hydroxy-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 20);-   N-1-((1S-2    syn)-1-Benzyl-2-hydroxy-3-(1-isobutyl-3,3-dimethylsulfonylurea)-propyl)-2-((quinoline-2-carbonyl)-amino)-succinamide    (compound 21);-   N-(4-(((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-isobutyl-sulfamoyl)-phenyl)-acetamide    (compound 48);-   N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-4-methoxy-benzenesulfonamide    (compound 100);-   4-Chloro-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 116);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 140);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(3-pyridin-3-yl-methoxycarbonylamino)-butyl-4-methoxy-benzenesulfonamide    (compound 142);-   N-Cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 148);-   N-Cyclohexylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-4-methoxy-benzenesulfonamide    (compound 158);-   N-(4-(Cyclohexylmethyl)-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-sulfamoyl-phenyl)-acetamide    (compound 160);-   N-cyclopentylmethyl-4-hydroxy-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-(pyridin-3-yl-methoxycarbonylamino)-butyl)-benzenesulfonamide    (compound 166);-   4-Amino-N-((2    syn,3S)-2-Hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 168);-   4-Amino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 171);-   2,4-Diamino-N-cyclopentylmethyl-N-((2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-benzenesulfonamide    (compound 173);-   4-Hydroxy-N-(2    syn,3S)-2-hydroxy-4-phenyl-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-isobutyl-benzenesulfonamide    (compound 175); and-   N-((2    syn,3S)-4-Cyclohexyl-2-hydroxy-3-((S)-tetrahydrofuran-3-yloxycarbonylamino)-butyl)-N-cyclopentylmethyl-4-methoxy-benzenesulfonamide    (compound 195).

The sulfonamides of this invention may be synthesized using conventionaltechniques. Advantageously, these compounds are conveniently synthesizedfrom readily available starting materials.

The compounds of this invention are among the most readily synthesizedHIV protease inhibitors known. Previously described HIV proteaseinhibitors often contain four or more chiral centers, numerous peptidelinkages and/or require air-sensitive reagents (such as organometalliccomplexes) to effect their synthesis. The relative ease with which thecompounds of this invention can be synthesized represents an enormousadvantage in the large scale production of these compounds.

In general, sulfonamides of formula I are conveniently obtained fromα-amino acid derivatives having the general formulaA-(B)_(x)—NH—CH(D)-COOH, wherein A, B, X and D are defined as above forthe compounds of formula I. Such α-amino acid derivatives are oftencommercially available or may be conveniently prepared from commerciallyavailable α-amino acid derivatives using known techniques. See, forexample, T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis”, 2nd Ed., John Wiley and Sons (1991). Although this inventionenvisions the use of racemic mixtures of such starting materials, whenx=0, a single enantiomer in the S configuration is preferred.

Using known techniques, the α-amino acid derivative of general formulaA-(B)_(x)—NH—CH(D)-COOH may be readily converted to an amino ketonederivative of general formula A-(B)_(x)—NH—CH(D)-CO—CH₂—X, wherein X isa leaving group which suitably activates the α-carbon (i.e., makes themethylene susceptible to nucleophilic attack). Suitable leaving groupsare well known in the art and include halides and sulfonates, such asmethanesulfonate, trifluoromethanesulfonate or 4-toluenesulfonate X mayalso be a hydroxyl which is converted in situ to a leaving group (e.g.by treatment with a trialkyl- or triarylphosphine in the presence of adialkylazodicarboxylate). Methods for the formation of such amino ketonederivatives also are well known to those of skill in the art (see, forexample, S. J. Fittkau, J. Prakt. Chem., 315, p. 1037 (1973)).Alternatively, certain amino ketone derivatives are commerciallyavailable (e.g., from Bachem Biosciences, Inc., Philadelphia, Pa.).

The amino ketone derivative may then be reduced to the correspondingamino alcohol, represented by the formulaA-(B)_(x)—NH—CH(D)-CH(OH)—CH₂—X. Many techniques for reduction of aminoketone derivatives such as A-(B)_(x)—NH—CH(D)-CO—CH₂—X are well known tothose of ordinary skill in the art (Larock, R. C. “Comprehensive OrganicTransformations”, pp. 527-547, VCH Publishers, Inc.© 1989 and referencescited therein). A preferred reducing agent is sodium borohydride. Thereduction reaction is conducted at a temperature of from about −40° C.to about 40° C. (preferably, at about 0° C. to about 20° C.), in asuitable solvent system such as, for example, aqueous or neattetrahydrofuran or a lower alcohol, such as methanol or ethanol.Although this invention envisions both stereospecific andnon-stereospecific reduction of the amino ketone derivativeA-(B)_(x)—NH—CH(D)-CO—CH₂—X, stereoselective reduction is preferred.Stereoselective reduction may be accomplished by use of chiral reagentsknown in the art. In the present invention, stereoselective reductionmay be conveniently achieved, for instance, under non-chelating reducingconditions, where chiral induction of the newly formed hydroxyl group isset by the stereochemistry of the D group (i.e., Felkin-Ahn addition ofhydride). We particularly prefer stereoselective reductions wherein theresulting hydroxyl is syn to D. We have found that when the hydroxylgroup is syn to D, the final sulfonamide product is an HIV proteaseinhibitor of higher potency than the anti diastereomer.

The hydroxyl group of the amino alcohol may optionally be protected byany known oxygen protecting group (such as trialkylsilyl, benzyl, oralkyloxymethyl) to yield a protected amino alcohol having the formulaA-(B)_(x)—NH—CH(D)-C(OR⁶)—CH₂—X, wherein R⁶ is H or any suitable hydroxyprotecting group. Several useful protecting groups are described in T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2dEd., John Wiley and Sons (1991).

The amino alcohol may then be reacted with a nucleophilic amine compoundto form an intermediate of formula III:

wherein D and R⁶ are as described above, and L is either D′ (asdescribed for compounds of formula I) or hydrogen.

In a particularly advantageous synthetic scheme, simultaneous activationof the methylene and protection of the alcohol may be accomplished byforming an N-protected amino epoxide from the oxygen and its adjacentmethylene to give an intermediate of formula II:

wherein A, B and D are as defined above for compounds of formula I.Suitable solvent systems for preparing the N-protected amino epoxideinclude ethanol, methanol, isopropanol, tetrahydrofuran, dioxane,dimethyl formamide and the like (including mixtures thereof). Suitablebases for producing the epoxide include alkali metal hydroxides,potassium t-butoxide, DBU and the like. A preferred base is potassiumhydroxide.

Reaction of the N-protected amino epoxide or other suitably activatedintermediates with an amine is carried out neat, i.e. in the absence ofsolvent, or in the presence of a polar solvent such as lower alkanols,water, dimethylformamide or dimethylsulfoxide. The reaction can becarried out conveniently between about 0° C. and 120° C., preferablybetween about 20° C. and 100° C. Alternatively, the reaction may becarried out in the presence of an activating agent, such as activatedalumina in an inert solvent, preferably an ether, such as diethyl ether,tetrahydrofuran, dioxane, or tert-butyl methyl ether, conveniently fromabout room temperature to about 110° C., as described by Posner andRogers, J. Am. Chem. Soc., 99, p. 8208 (1977). Other activating reagentsinclude lower trialkylaluminum species, such as triethylaluminum, ordialkylaluminum halide species, such as diethylaluminum chloride(Overman and Flippin, Tetrahedron Letters, p. 195 (1981)). Reactionsinvolving these species are conveniently carried out in inert solventssuch as dichloromethane, 1,2-dichloroethane, toluene, or acetonitrilebetween about 0° C. and about 110° C. Further methods of displacingleaving groups, or opening epoxides with amines or their equivalentssuch as azides or timethylsilyl cyanide (Gassman and Guggenheim, J. Am.Chem. Soc. 104, p. 5849 (1982)), are known and will be apparent to thoseof ordinary skill in the art.

Compounds of formulae II and III, and functionality-protectedderivatives thereof, are useful as intermediates for the preparation ofcompounds of formula I. In those cases where L represents D′, compoundsof formula III may be converted to compounds of formula I by reactionwith sulfonyl-activated species to form sulfonamides, sulfonyl ureas,thiocarbamates and the like. Methods for preparing suchsulfonyl-activated species are well within the ordinary skill of theart. Typically, sulfonyl halides are used to obtain sulfonamides. Manysulfonyl halides are commercially available; others may be easilyobtained using conventional synthetic techniques (Gilbert, E. E. “RecentDevelopments in Preparative Sulfonation and Sulfation” Synthesis 1969: 3(1969) and references cited therein; Hoffman, R. V.“M-Trifluoromethylbenzenesulfonyl Chloride” Org. Synth. Coll. Vol. VII,John Wiley and Sons (1990); Hartman, G. D. et. al. “4-SubstitutedThiophene- and Furan-2-sulfonamides as Topical Carbonic AnhydraseInhibitors” J. Med. Chem., 35, p. 3822 (1992) and references citedtherein. Sulfonyl ureas are usually obtained by the reaction of an aminewith sulfuryl chloride or a suitable equivalent such assulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.Thiocarbamates are typically obtained by the reaction of an alcohol withsulfuryl chloride or a suitable equivalent such assulfuryl-bis-imidazole or sulfuryl-bis-N-methyl imidazole.

In the case of compounds of formula III wherein L is hydrogen,conversion of the resultant primary amine to a secondary amine may becarried out by known techniques. Such techniques include reaction withan alkyl halide of alkyl sulfonate, or by reductive alkylation with analdehyde or carboxylic acid or activated derivative thereof using, forinstance, catalytic hydrogenation or sodium cyanoborohydride (Borch etal., J. Am. Chem. Soc., 93, p. 2897 (1971)). Alternatively, the primaryamine may be acylated followed by reduction with borane or anothersuitable reducing reagent, for example, as described by Cushman et al.,J. Org. Chem., 56, p. 4161 (1991). This technique is especially usefulin compounds of formula III where B is absent and A represents aprotecting group such as tert-butoxycarbonyl (Boc) or benzyloxycarbonyl(Cbz).

If variable A of a particular compound of formula I represents aremovable protecting group, removal of that group followed by reactionof the resulting amine with an appropriate activated reagent willadvantageously yield a different compound of formula I. For instance,reaction with an activated carboxylate, such as an acyl halide (e.g.,acid fluo-rides, acid chlorides, and acid bromides), an activated estersuch as nitrophenyl ester or 1-hydroxysuccinimide (HOSu) ester, ananhydride such as the symmetrical anhydride or isobutyl anhydride, ormixed carbonic-phosphoric or carbonic-phosphinic anhydrides, will yieldthe corresponding amide. Ureas may be obtained by reaction withisocyanates or amines in the presence of bis-activated carbonic acidderivatives such as phosgene or carbonyldiimdazole. Carbamates may beobtained by reaction with chlorocarbonates, with carbonates esterifiedwith leaving groups such as 1-hydroxybenzotriazole (HOBT) or HOSu, orwith alcohols in the presence of bis-activated carbonic acid derivativessuch as phosgene or carbonyldiimdazole. It will be readily recognizedthat in order to facilitate specific reactions, the protection of one ormore potentially reactive groups followed by subsequent removal of thatgroup may be required. Such modification to the reaction schemesoutlined above are within the ordinary skill of the art.

If variable B of a particular compound of formula I is absent andvariable A of that compound represents a removable protecting group,removal of A, followed by reaction of the resulting amine with an aminoacid or suitably N-protected derivative thereof, followed by asubsequent reaction of the free α-amine if present, as described above,will yield a further compound of formula I. The addition of amino acidsand their derivatives is accomplished by well known methods of peptidesynthesis. Some of these methods are generally set forth in Bodanszkyand Bodanszky, “The Practice of Peptide Synthesis”, Springer-Verlag,Berlin, Germany (1984) and in the “The Peptides”, Gross and Meinhofer(Eds); Academic Press, 1979, Vols. I-III, which are incorporated hereinby reference.

Typically, for solution phase synthesis of peptides, the α-amine of theamino acid to be coupled is protected by Boc, Cbz, allyloxycarbonyl(Alloc) or 9-fluorenylmethoxycarbonyl (Fmoc), while the free carboxyl isactivated by reaction with a carbodiimide such asdicyclohexylcarbodiimide (DCC),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), ordiisopropylcarbodiimide (DIC), optionally in the presence of a catalystsuch as HOBT, HOSu, or dimethylaminopyridine (DMAP). Other methods whichproceed through the intermediacy of activated esters, acid halides,enzyme-activated amino acids and anhydrides includingN-carboxy-anhydrides, symmetrical anhydrides, mixed carbonic anhydrides,carbonic-phosphinic and carbonic-phosphoric anhydrides are alsosuitable. After the peptide has been formed, protecting groups may beremoved by methods described in the references listed above, such as byhydrogenation in the presence of a palladium, platinum or rhodiumcatalyst, treatment with sodium in liquid ammonia, hydrochloric,hydrofluoric, hydrobromic, formic, trifluoromethanesulfonic, ortrifluoroacetic acid, secondary amines, fluoride ion, trimethylsilylhalides including bromide and iodide, or alkali.

One particularly useful synthetic scheme for producing sulfonamides offormula XV is shown below:

Compounds of formula X may be advantageously synthesized from readilyavailable starting materials (see D. P. Getman, J. Med. Chem., 36, p.288 (1993)). Each step of the above synthetic scheme may be carried outas generally described above.

A particularly useful synthetic scheme for producing the preferredsulfonamides of formula XXII is shown below:

Compounds of formula XX may be advantageously synthesized from readilyavailable starting materials (see B. E. Evans et al., J. Org. Chem., 50,p. 4615 (1985)). Each step of the above synthetic scheme may be carriedout as generally described above.

After converting a compound of formula XX to a compound of formula XXI,as detailed in the previous reaction scheme, the compound of formula XXImay alternatively be reacted with an amino acid or amino acidderivative, as described generally above, to yield a preferred compoundof formula XXXI. A particularly useful synthetic scheme utilizing thisstrategy is set forth below:

As can be appreciated by the skilled artisan, the above syntheticschemes are not intended to comprise a comprehensive list of all meansby which the compounds described and claimed in this application may besynthesized. Further methods will be evident to those of ordinary skillin the art.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds of formula I are characterized by a superior ability toinhibit HIV protease activity and viral replication. We believe thatthis is due to specific steric and electronic interactions between theprotease and compounds of formula I. This belief stems from our analysisof the structural basis for the activity of compounds of formula I, inview of the known crystal structures of HIV protease and boundinhibitors, such as the structure reported in Miller et al. “Structureof Complex of Synthetic HIV-1 Protease with a Substrate-Based Inhibitorat 2.3 Å Resolution”, Science, vol. 246, pp. 1149-1152 (1989), which isincorporated herein by reference, as well as structures determined inour laboratories. According to these structures, the active site of HIVaspartyl protease is defined by a deep groove containing subpockets foraccommodation of various side chains of the protease substrate—referredto as P₁-P_(n) and P₁′-P_(n)′, according to conventional proteasenomenclature. In the center of the groove, lie two aspartic acidresidues (Asp25 and Asp25′ according to the numbering system of Milleret al.) in a manner typical of the active site aspartates of knownaspartyl proteases, which are believed to be the catalytic residues ofthe enzyme. The groove is covered by two C₂-symmetrically disposed“flaps” which also make various direct and indirect contacts with boundsubstrates.

We believe that the substituents A, D, D′ and E of the compounds offormula I associate with HIV protease by way of hydrophobic forces inthe binding pockets of the enzyme. We also believe that the sulfonamidegroup hydrogen binds tightly to a water molecule held by hydrogen bondsto the flaps of the protease (“the flap water molecule”; water molecule511, according to the Miller et al. numbering system).

In view of the above discovery, an alternative embodiment of thisinvention relates to novel HIV protease inhibitors possessing certainstructural and physicochemical features. We have discovered thatcompounds possessing the following novel combination of features aresurprisingly effective HIV protease inhibitors:

(1) a first and a second hydrogen bond acceptor moiety, at least one ofwhich is more highly polarizable than a carbonyl, said moieties beingthe same or different, and being capable of hydrogen bonding with thehydrogen atoms of the flap water molecule of an HIV aspartyl proteasewhen the compound is bound thereto;

(2) substantially hydrophobic moieties which associate with the P₁ andP₁′ binding pockets of said HIV aspartyl protease when the compound isbound thereto;

(3) a third hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease when the compound is boundthereto;

(4) an additional occupied volume of space of at least 100 Å³ when thecompound is bound to the active site of said HIV aspartyl protease, saidspace overlapping with the volume of space that would be filled by anative substrate of said HIV aspartyl protease or a nonhyrolyzableisostere thereof;

(5) a deformation energy of binding of the compound to said HIV aspartylprotease of not greater than 10 kcal/mole; and

(6) a neutral or favorable enthalpic contribution from the sum of allelectrostatic-interactions between the compound and the protease whenthe compound is bound to said HIV aspartyl protease.

Compounds having the above-cited features can be readily identified ordesigned by one of ordinary skill in the art using a combination ofchemical reasoning and computational methods. For example, those ofordinary skill in the art can readily identify or choose hydrogenbonding and hydrophobic moieties or groups required in features (1)-(3),while features (4)-(6) can be ascertained using well known computationalmethods for determination of structural (e.g. conformational) andenergetic properties of molecules.

Furthermore, compounds characterized by features (1) through (6) listedabove may be obtained using any conventional technique, includingchemical synthesis and natural product isolation. We prefer using thesynthetic schemes detailed above for compounds of formula I.

We have discovered that when an HIV protease inhibitor forms hydrogenbonds to the flap water molecule through two hydrogen bonding moieties,at least one of which is more highly polarizable than a carbonyl, theability of those compounds to inhibit HIV protease activity isdramatically improved, as compared with conventional HIV proteaseinhibitors.

While not wishing to be bound by theory, we believe that the stronghydrogen bonds that form between the flap water molecule and the twohydrogen bonding moieties, at least one of which is more highlypolarizable than a carbonyl, lower the overall binding energy of theinhibitor. Most HIV protease inhibitors known in the art utilize onlycarbonyl groups for hydrogen bonding to the flap water molecule and are,thus, inferior to those of the present invention. We believe that theincreased polarization that results from the large dipole moment of thehighly polarizable hydrogen bonding moiety (as compared to the dipolemoment of a carbonyl moiety) creates a stronger and tighter hydrogenbond with the flap water molecule. We prefer to utilize tetravalentoxygenated sulfur, hexavalent oxygenated sulfur and pentavalentoxygenated phosphorus as the highly polarizable hydrogen bonding moiety.Tetravalent oxygenated sulfur and hexavalent oxygenated sulfur are morepreferred as the highly polarizable hydrogen bonding moiety. Hexavalentoxygenated sulfur (—SO₂—) is most preferred.

We have found that when the highly polarizable hydrogen bonding moietyis a sulfonamide, the overall binding energy of the inhibitor isparticularly low. We believe that this increased stability is due toparticular conformational characteristics of the sulfonamide S—N bond.Specifically, the sulfonamide S—N bond exists in only two low-energyrotamers (see J. B. Nicholas et al., J. Phys. Chem., 95, p. 9803 (1991)and R. D. Bindal et al., J. Am. Chem. Soc., 112, p. 7861 (1990)). Thishas the effect of locking that portion of the molecule into a favorableconformation wherein one or both of the highly polarized S═O oxygens canbe involved in hydrogen bonding interactions with the flap water.

The remaining five structural and physicochemical features recited above(i.e., features (2) through (6)) are generally recognized in the art toimprove the ability of a compound to competitively inhibit HIV proteaseactivity. Although there are several other features thought to increasethe inhibitory property (such as binding of the inhibitor backbone tothe enzyme), we have discovered that the combination of the fiveabove-cited elements alone, together with novel element (1), typifieseffective HIV protease inhibitors.

In general, the binding energy of a particular protease inhibitor islowered when hydrophobic moieties on the inhibitor are located so as toassociate with the enzyme's hydrophobic binding pockets. In the case ofHIV-1 protease, the location and nature of the P₁ and P₁′ bindingpockets are known to those of ordinary skill in the art (see, forexample, M. Miller et al., cited above). Substantially hydrophobic sidechains which fit into the well defined P₁ and P₁′ binding pockets arealso known to those in the art. Preferred side chains are located within4 Å of the enzyme when bound to HIV protease. Preferred hydrophobic sidechains include those substantially similar to those of hydrophobicnatural and unnatural α-amino acids, including alanine, valine, leucine,isoleucine, methionine, phenylalanine, α-amino isobutyric acid,alloisoleucine, tyrosine, and tryptophan. Insofar as a portion of thisside chain is in contact with bulk solvent or protrudes out of theenzyme, it is not considered to be wholly within P₁ or P₁′ and maycontain polar functionality such as a charged amine at that location.

It has also been established in the art that the presence of a hydroxylgroup within hydrogen bond proximity to the two catalytic aspartic acidresidues of HIV protease (Asp25 and Asp25′) is an important feature ofan effective HIV protease inhibitor (see, for example, R. Bone et al.,“X-ray Crystal Structure of the HIV Protease Complex with L-700,417, anInhibitor with Pseudo C₂ Symmetry”, J. Am. Chem. Soc., 113, pp. 9382-84(1991)). It is further understood that the geometry of the Asp-bindinghydrogen bonding moiety is of particular importance. Although we preferto use a hydroxyl group at this position, any hydrogen bonding moietythat is capable of forming hydrogen bonds with the Asp residues isacceptable. Such hydrogen bonding moieties are known to those of skillin the art (e.g., phosphinic acid (D. Grobelny et al., Biochem. Biophys.Res. Commun., 169, p. 1111 (1990)).

It is further understood that binding of competitive inhibitors to HIVprotease is optimally accomplished by having the inhibitor traverse avolume overlapping that occupied by the native polypeptide substratewhen it is bound to the active site of the enzyme. Effective HIVprotease inhibitors typically have a relatively small difference inenergy between their bound and free states (i.e., a small deformationenergy of binding). The most preferred HIV protease inhibitors of thisinvention have a deformation energy of binding of not greater than 10kcal/mole (preferably, not greater than 7 kcal/mole). It should benoted, however, that HIV protease inhibitors may interact with HIVprotease in more than one conformation which is similar in overallbinding energy (see K. H. M. Murthy, J. Biol. Chem., 267, (1992)). Inthose cases, the deformation energy of binding is taken to be thedifference between the energy of the free compound and the averageenergy of the conformations observed when the inhibitor binds to theenzyme.

Furthermore, it is understood that the most effective proteaseinhibitors also lack repulsive electrostatic interaction with the targetprotease in their bound state. Such non-complementary (e.g.,electrostatic) interactions include repulsive charge-charge,dipole-dipole and charge-dipole interactions. Specifically, in the mostpreferred HIV protease inhibitors of this invention, the sum of allelectrostatic interactions between the compound and the enzyme when thecompound is bound to HIV protease makes a neutral or favorablecontribution to the enthalpy of binding.

Preferred compounds characterized by the above features (1)-(6) arecompounds of formula XL:

Z¹-Q¹-L¹-M-L²-Q²-Z²  (XL)

wherein:

Q¹ and Q² are independently hydrogen bond acceptor moieties capable ofbinding with the hydrogen atoms of the flap water molecule of an HIVaspartyl protease, with the proviso that at least one of Q¹ or Q² ismore highly polarizable than a carbonyl;

M is a hydrogen bonding moiety, which may be either a hydrogen bonddonor or acceptor, capable of simultaneously hydrogen bonding to Asp25and Asp25′ of said HIV aspartyl protease;

L¹ and L² are independently acyclic or cyclic linker moieties; and

each of Z¹ and Z² may be optionally present and, if present, areindependently selected from groups which occupy a volume of spaceoverlapping with the volume of space that would be filled by the nativesubstrate of said HIV aspartyl protease.

More preferred compounds of formula XL contain at least one group Q¹ orQ² comprising —SO₂—. Most preferred compounds of formula XL contain atleast one group Q¹ or Q² comprising a substituted sulfonamide.

In one embodiment of this invention, compounds of formula XL may befurther constrained by “conformational locks”, such as a macrocyclicring structure. Such constraints are well known in the art ofpeptidomimetics and may result in compounds with strong biologicalactivity. See, for example, Dhanoa, D. S. et al. “The Synthesis ofPotent Macrocyclic Renin Inhibitors” Tetrahedron Lett. 33, 1725 (1992)and Flynn, G. A. et al. “An Acyl-Iminium Ion Cyclization Route to aNovel Conformationally Restricted Dipeptide Mimic: Applications toAngiotensin-Converting Enzyme Inhibition” J. Am. Chem. Soc. 109, 7914(1989)).

This invention also includes novel methods for accurate identification,design, or prediction of HIV inhibitors characterized by structural andphysicochemical features (1) through (6). By virtue of these methods,the skilled artisan can routinely predict and produce particularlyeffective HIV protease inhibitors.

We have found that the following method for identification, design orprediction of effective HIV protease inhibitors is particularly useful;

-   -   (a) selecting a candidate compound of defined chemical structure        containing a first and a second hydrogen bond acceptor moiety,        at least one of which is more highly polarizable than a        carbonyl, said moieties being the same or different; a third        hydrogen bonding moiety, which may be either a hydrogen bond        donor or acceptor; and at least two substantially hydrophobic        moieties;    -   (b) determining a low-energy conformation for binding of said        compound to the active site of an HIV aspartyl protease;    -   (c) evaluating the capability of said first and second hydrogen        bond acceptor moieties to form hydrogen bonds to the flap water        molecule of said HIV aspartyl protease when said compound is        bound thereto in said conformation;    -   (d) evaluating the capability of said substantially hydrophobic        moieties to associate with the P₁ and P₁′ binding pockets of        said HIV aspartyl protease when said compound is bound thereto        in said conformation;    -   (e) evaluating the capability of said third hydrogen bonding        moiety to form hydrogen bonds to Asp25 and Asp2S′ of said HIV        aspartyl protease when said compound is bound thereto in said        conformation;    -   (f) evaluating the overlap of the occupied volume of said        compound when said compound is bound to said HIV aspartyl        protease in said conformation and the occupied volume of a        native substrate of HIV aspartyl protease or a nonhydrolyzable        isostere thereof, when said polypeptide is bound to said HIV        aspartyl protease;    -   (g) evaluating the deformation energy of binding of said        compound to said HIV aspartyl protease;    -   (h) evaluating the enthalpic contribution of the sum of all        electrostatic interactions between said compound and said HIV        aspartyl protease when said compound is bound thereto in said        conformation; and    -   (i) accepting or rejecting said candidate compound as an HIV        protease inhibitor based upon the determinations and evaluations        carried out in steps (b) through (h).

Using the novel combination of steps set forth in this screening method,the skilled artisan can advantageously avoid time consuming andexpensive experimentation to determine enzymatic inhibition activity ofparticular compounds. The method is also useful for facilitatingrational design of HIV protease inhibitors and anti-HIV viral agents,including therapeutic and prophylactic agents against HIV infection.Accordingly, the present invention relates to such inhibitors andanti-viral agents produced by the screening method described above.

A variety of conventional techniques may be used to carry out each ofthe above evaluations. Generally, these techniques involve determiningthe location and binding proximity of a given moiety, the occupiedvolume of space of a bound compound, the deformation energy of bindingof a given compound and electrostatic interaction energies. Examples ofconventional techniques useful in the above evaluations include: quantummechanics, molecular mechanics, molecular dynamics, Monte Carlosampling, systematic searches and distance geometry methods (G. R.Marshall, Ann. Ref. Pharmacol. Toxicol., 27, p. 193 (1987)). Specificcomputer software has been developed for use in carrying out thesemethods. Examples of programs designed for such uses include: Gaussian92, revision C (M. J. Frisch, Gaussian, Inc., Pittsburgh, Pa. ©1992);AMBER, version 3.0 (U.C. Singh, University of California at SanFrancisco, ©1992); QUANTA/CHARMM (Molecular Simulations, Inc.,Burlington, Mass. ©1992); and Insight II/Discover (Biosysm TechnologiesInc., San Diego, Calif. ©1992). These programs may be implemented, forinstance, using a Silicon Graphics workstation, IRIS 4D/35 or IBMRISC/6000 workstation model 550. Other hardware systems and softwarepackages will be known and of evident applicability to those skilled inthe art.

Additional analysis of the actual detailed interactions of the HIVprotease-inhibitor complex can be employed to ascertain morespecifically the binding associations between the enzyme and the boundinhibitor. Such analysis may be carried out, for example, by studying asolution of the complex by single- and multi-dimensional NMR techniques.Advantageously, the enzyme and/or the inhibitor may be enriched withstable isotopes such as ¹³C, ¹⁵N and ²H to more easily determine bindingconformation and proximity. Techniques, such as isotope editing, may beused to enhance the resolution with which the interactions are observed.

Either as an alternative or a supplemental analysis, the HIVprotease-inhibitor complex may be studied by single crystal X-raydiffraction. The process of determining the structures ofprotein/inhibitor complexes using the X-ray techniques described aboveis well known and has been used for many different complexes (see T. L.Blundel and L. N. Johnson, Protein Crystallography, Academic Press,(1976) and Methods in Enzymology, volumes 114 and 115, H. W. Wyckoff etal., eds., Academic Press (1985)). This technique can employ, forinstance, a highly purified preparation of HIV protease complexed withan inhibitor of interest in a buffered solution (typically at a pH ofbetween about 4.5 and about 8.0). The complex is allowed to crystallizein the presence of a precipitation agent (such as ammonium sulfate)under conditions which yield single crystals of the complex. Specificconditions for crystallizing HIV protease with various inhibitors havebeen well documented (see, for example, G. B. Dreyer et al.,Biochemistry, 31, p. 6646 (1992)). Application of a concentrated X-raybeam to an appropriately prepared and mounted crystal (preferably, anX-ray beam from a rotating anode X-ray generator or synchrotron) willyield a diffraction pattern from the reflected X-ray beam.

Detection of the diffracted rays may be carried out by visualizingphotographic paper exposed to the diffracted X-rays or alternatively, byusing a multiwire area detector (such as that manufactured by SiemensAnalytical X-Ray Instruments, Inc. (Madison, Wis.)) or an R-axis IIimage plate system from Rigaku Corporation (distributed by MolecularStructure Corporation, The Woodlands, Tex.). Other systems forgenerating and collecting X-ray diffraction data will be known to thoseof ordinary skill in the art.

Refinement of the X-ray diffraction data yields a three dimensionalstructure. Computer software (such as X-PLOR (Yale University, ©1992,distributed by Molecular Simulations, Inc.) has been developed to carryout this refinement.

In general, using the above techniques with an appropriately preparedcrystalline complex, a structure may be refined to about 2-3 Å with an Rvalue of about 0.25 or less. As the skilled artisan can appreciate,these values are adequate to determine the interactions between HIVprotease and a given compound such that it will be clear if features (1)through (6) are present and consequently, whether that given compound isan HIV aspartyl protease inhibitor. Thus, additional inhibitorsaccording to this invention may be designed and predicted based on acombination of crystallographic structural information and computationalanalysis.

For example, to predict the binding of a candidate inhibitor accordingto this invention, the inhibitor is examined to determine whether themolecule contains functionality which is not well represented by theexisting forcefield models in CHARMM (Molecular SimulationsIncorporated, Burlington, Mass.) or AMBER (Professor P. A. Kollman,UCSF). If any functionality is not well represented, we then examine allpublished structural information for molecules containing suchfunctionality, and in some cases perform high-level ab initiocalculations on simple molecules containing these functionalities todetermine their preferred conformations and the energy differencesbetween various conformations. More accurate parameters describing thesefunctional groups may then be derived for the CHARMM and/or AMBERforcefields and used in subsequent calculations.

Next, the candidate inhibitor is aligned in 3-dimensional space withother, related inhibitors whose bound conformations have previously beendetermined by x-ray crystallography. Both Van der Walls volume andelectrostatic potentials are used to direct the alignment process. Thealignment is typically done with software like Quanta (MolecularSimulations) or InsightII (Biosym Technologies, San Diego, Calif.). Thisalignment can be done manually within this software, or more automatedalignment procedures within the software (e.g. the “superimpose” optionof Quanta or the “APEX” module of InsightII) may be used. The result ofthis alignment is a first guess of the “bound” conformation of thecandidate inhibitor. This inhibitor is then docked in the active site ofHIV protease, and the conformation is energy minimized with the enzymeatoms held fixed in space. These minimizations are typically done usingthe CHARMM or AMBER forcefields.

Because inhibitors can sometimes bind in multiple or unexpectedconformations within an active site, we often then carry out furthersearches of the bound conformation of the enzyme-inhibitor complex. Forexample, a variety of Monte Carlo search techniques (e.g. as found inthe Conformational Search Module of Quanta) may be used, along withhigh-temperature dynamics and simulated annealing. These searchtechniques reveal whether there are alternative, reasonable low-energyconformations in which the inhibitor may bind to the enzyme. The effectsof solvation and desolation in the formation of the variousenzyme-inhibitor complexes may be estimated with programs such as DELPHI(Biosym), Polaris (Molecular Simulations) and AMSOL (Professor C.Cramer, University of Minnesota). The result of this searching is a setof one or more bound conformations for the candidate inhibitor.

For each of the low-energy conformations, waters may then be added tothe active site of the enzyme and the entire system relaxed. Finally,molecular dynamics simulations may be used to study the detailed motionsof the enzyme, the inhibitor, and related water molecules.

The final set of remaining low-energy conformations (typically a verysmall number) represents our predictions of the bound conformation ofthe candidate inhibitor. Each conformation includes our estimate of thedynamic flexibility of the entire system (inhibitor, enzyme, andwaters).

The more advanced methodology is typically applied to the study of thefirst few compounds in a series, when there are the greatestuncertainties about the possible binding mode(s) in the enzyme activesite. For later compounds within a series, the low energy conformersobtained from the searches on earlier compounds provide informationabout the possible low energy conformers of the inhibitor compounds. Inaddition, crystallographic information about the conformation of thebound complexes of earlier compounds within a series is often available.This prior computational and structural work advantageously facilitatesthe prediction of the bound conformation of candidate inhibitormolecules.

To exemplify the above screening method, we have carried out thefollowing evaluation of compound 140 (Table II), a preferred compound ofthis invention, as described below.

Prediction of Binding Conformation and Energy of Compound 140 to HIVProtease

The forcefield for the benzenesulfonamide portion of compound 140 wasderived from ab initio calculations and incorporated into the AMBERforcefield. The latest CHARMM forcefield parameters for this moiety werefound to be adequate for energy minimization studies and are used in allQuanta/CHARMM calculations.

The low energy conformers obtained from the conformational searches onearlier compounds in the sulfonamide series (such as compound 16)provided information about the possible low energy conformers ofcompound 140. These low energy conformers were aligned in 3-dimensionalspace with other related inhibitors whose bound conformations havepreviously been determined by x-ray crystallography. This alignmentprocess was carried out manually within Quanta and, in some cases, wasassisted with the “conformational search” option of Quanta. Thereference crystal structure used in this alignment was the complex ofHIV-1 protease with compound 16. This inhibitor structure was energyminimized in the active site of the enzyme using Quanta/CHARM. Theenzyme atoms were held fixed during this minimization. Only the flapwater was included. Later simulations allowed the enzyme to relax andused a variety of dielectric approximations. A single low-energyconformation which was consistent with all previous conformationalsimulations and crystallographic data was obtained (see FIG. 1). Thispredicted binding conformation was later found to be essentially inagreement with the results obtained by x-ray crystallography (see FIGS.2 and 3).

As discussed above, the novel compounds of the present invention areexcellent ligands for aspartyl proteases, particularly HIV-1 and HIV-2proteases. Accordingly, these compounds are capable of targeting andinhibiting late stage events in HIV replication, i.e., the processing ofthe viral polyproteins by HIV encoded proteases. Such compounds inhibitthe proteolytic processing of viral polyprotein precursors by inhibitingaspartyl protease. Because aspartyl protease is essential for theproduction of mature virions, inhibition of that processing effectivelyblocks the spread of virus by inhibiting the production of infectiousvirions, particularly from chronically infected cells. Compoundsaccording to this invention advantageously inhibit the ability of theHIV-1 virus to infect immortalized human T cells over a period of days,as determined by an assay of extracellular p24 antigen—a specific markerof viral replication. Other anti-viral assays have confirmed the potencyof these compounds.

The compounds of this invention may be employed in a conventional mannerfor the treatment of viruses, such as HIV and HTLV, which depend onaspartyl proteases for obligatory events in their life cycle. Suchmethods of treatment, their dosage levels and requirements may beselected by those of ordinary skill in the art from available methodsand techniques. For example, a compound of this invention may becombined with a pharmaceutically acceptable adjuvant for administrationto a virally-infected patient in a pharmaceutically acceptable mannerand in an amount effective to lessen the severity of the viralinfection.

Alternatively, the compounds of this invention may be used in vaccinesand methods for protecting individuals against viral infection over anextended period of time. The compounds may be employed in such vaccineseither alone or together with other compounds of this invention in amanner consistent with the conventional utilization of proteaseinhibitors in vaccines. For example, a compound of this invention may becombined with pharmaceutically acceptable adjuvants conventionallyemployed in vaccines and administered in prophylactically effectiveamounts to protect individuals over an extended period time against HIVinfection. As such, the novel protease inhibitors of this invention canbe administered as agents for treating or preventing HIV infection in amammal.

The compounds of formula I, especially those having a molecular weightof less than about 700 g/mole, may be readily absorbed by thebloodstream of mammals upon oral administration. Compounds of formula Ihaving a molecular weight of less than about 600 g/mole are most likelyto demonstrate oral availability. This surprisingly impressive oralavailability makes such compounds excellent agents fororally-administered treatment and prevention regimens against HIVinfection.

The compounds of this invention may be administered to a healthy orHIV-infected patient either as a single agent or in combination withother anti-viral agents which interfere with the replication cycle ofHIV. By administering the compounds of this invention with otheranti-viral agents which target different events in the viral life cycle,the therapeutic effect of these compounds is potentiated. For instance,the co-administered anti-viral agent can be one which targets earlyevents in the life cycle of the virus, such as cell entry, reversetranscription and viral DNA integration into cellular DNA. Anti-HIVagents targeting such early life cycle events include, didanosine (ddI),alcitabine (ddC), d4T, zidovudine (AZT), polysulfated polysaccharides,sT4 (soluble CD4), ganiclovir, dideoxycytidine, trisodiumphosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferonand trimenotrexate. Additionally, non-nucleoside inhibitors of reversetranscriptase, such as TIBO or nevirapine, may be used to potentiate theeffect of the compounds of this invention, as may viral uncoatinginhibitors, inhibitors of trans-activating proteins such as tat or rev,or inhibitors of the viral integrase.

Combination therapies according to this invention exert a synergisticeffect in inhibiting HIV replication because each component agent of thecombination acts on a different site of HIV replication. The use of suchcombinations also advantageously reduces the dosage of a givenconventional anti-retroviral agent which would be required for a desiredtherapeutic or prophylactic effect as compared to when that agent isadministered as a monotherapy. These combinations may reduce oreliminate the side effects of conventional single anti-retroviral agenttherapies while not interfering with the anti-retroviral activity ofthose agents. These combinations reduce potential of resistance tosingle agent therapies, while minimizing any associated toxicity. Thesecombinations may also increase the efficacy of the conventional agentwithout increasing the associated toxicity. In particular, we havediscovered that these compounds act synergistically in preventing thereplication of HIV in human T cells. Preferred combination therapiesinclude the administration of a compound of this invention with AZT,ddI, ddC or d4T.

Alternatively, the compounds of this invention may also beco-administered with other HIV protease inhibitors such as Ro 31-8959(Roche), L-735,524 (Merck), XM 323 (Du-Pont Merck) and A-80,987 (Abbott)to increase the effect of therapy or prophylaxis against various viralmutants or members of other HIV quasi species.

We prefer administering the compounds of this invention as single agentsor in combination with retroviral reverse transcriptase inhibitors, suchas derivatives of AZT, or other HIV aspartyl protease inhibitors. Webelieve that the co-administration of the compounds of this inventionwith retroviral reverse transcriptase inhibitors or HIV aspartylprotease inhibitors may exert a substantial synergistic effect, therebypreventing, substantially reducing, or completely eliminating viralinfectivity and its associated symptoms.

The compounds of this invention can also be administered in combinationwith immunomodulators (e.g., bropirimine, anti-human alpha interferonantibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha,diethyldithiocarbamate, tumor necrosis factor, naltrexone and rEPO); andantibiotics (e.g., pentamidine isethiorate) to prevent or combatinfection and disease associated with HIV infections, such as AIDS andARC.

When the compounds of this invention are administered in combinationtherapies with other agents, they may be administered sequentially orconcurrently to the patient. Alternatively, pharmaceutical orprophylactic compositions according to this invention may be comprisedof a combination of an aspartyl protease inhibitor of this invention andanother therapeutic or prophylactic agent.

Although this invention focuses on the use of the compounds disclosedherein for preventing and treating HIV infection, the compounds of thisinvention can also be used as inhibitory agents for other viruses whichdepend on similar aspartyl proteases for obligatory events in their lifecycle. These viruses include, as well as other AIDS-like diseases causedby retroviruses, such as simian immunodeficiency viruses, but are notlimited to, HTLV-I and HTLV-II. In addition, the compounds of thisinvention may also be used to inhibit other aspartyl proteases, and inparticular, other human aspartyl proteases, including renin and aspartylproteases that process endothelin precursors.

Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle. Pharmaceutically acceptable carriers, adjuvants and vehiclesthat may be used in the pharmaceutical compositions of this inventioninclude, but are not limited to, ion exchangers, alumina, aluminumstearate, lecithin, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. We prefer oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. The termparenteral as used herein includes subcutaneous, intracutaneous,intravenous, intramuscular, intra-articular, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant such as Ph. Helv or a similar alcohol.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carriers which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in a capsuleform, useful diluents include lactose and dried corn starch. Whenaqueous suspensions are administered orally, the active ingredient iscombined with emulsifying and suspending agents. If desired, certainsweetening and/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible by topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxy-ethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-transdermal patches arealso included in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

Dosage levels of between about 0.01 and about 100 mg/kg body weight perday, preferably 0.5 and about 50 mg/kg body weight per day of the activeingredient compound are useful in the prevention and treatment of viralinfection, including HIV infection. Typically, the pharmaceuticalcompositions of this invention will be administered from about 1 toabout 5 times per day or alternatively, as a continuous infusion. Suchadministration can be used as a chronic or acute therapy. The amount ofactive ingredient that may be combined with the carrier materials toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Preferably,such preparations contain from about 20% to about 80% active compound.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level, treatment should cease.Patients may, however, require intermittent treatment on a long-termbasis upon any recurrence of disease symptoms.

As the skilled artisan will appreciate, lower or higher doses than thoserecited above may be required. Specific dosage and treatment regimensfor any particular patient will depend upon a variety of factors,including the activity of the specific compound employed, the age, bodyweight, general health status, sex, diet, time of administration, rateof excretion, drug combination, the severity and course of theinfection, the patient's disposition to the infection and the judgmentof the treating physician.

The compounds of this invention are also useful as commercial reagentswhich effectively bind to aspartyl proteases, particularly HIV aspartylprotease. As commercial reagents, the compounds of this invention, andtheir derivatives, may be used to block proteolysis of a target peptideor may be derivatized to bind to a stable resin as a tethered substratefor affinity chromatography applications. These and other uses whichcharacterize commercial aspartyl protease inhibitors will be evident tothose of ordinary skill in the art.

In order that this invention be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

General Materials and Methods

All temperatures are recorded in degrees Celsius. Thin layerchromatography (TLC) was carried out using 0.25 mm thick E. Merck silicagel 60 F₂₅₄ plates and elution with the indicated solvent system.Detection of the compounds was carried out by treating the plate with anappropriate visualizing agent, such as 10% solution of phosphomolybdicacid in ethanol or a 0.1% solution of ninhydrin in ethanol, followed byheating, and/or by exposure to UV light or iodine vapors whenappropriate. Thick layer silica gel chromatography was also carried outusing E. Merck 60 F₂₅₄ plates (“prep plates”) of 0.5, 1.0, or 2.0 mmthickness. Following development of the plate, the band of silicacontaining the desired compound was isolated and eluted with anappropriate solvent. Analytical HPLC was carried out using a Water'sDelta Pak, 5 μM silica, C18 reversed-phase column, 3.9 mm ID×15 cm Lwith a flow rate of 1.5 mL/min using the following table:

-   -   Mobile phase: A=0.1% CF₃CO₂H in H₂O        -   B=0.1% CF₃CO₂H in CH₃CN    -   Gradient: T=0 min., A (95%), B (5%)        -   T=20 min., A (0%), B (100%)        -   T=22.5 min., A (0%), B (100%)            Preparative HPLC was also carried out using C₁₈            reversed-phase media. HPLC retention times were recorded in            minutes. NMR spectral data was recorded using a Bruker            AMX500, equipped with either a reverse or QNP probe, at 500            MHz, and was taken in the indicated solvent.

We have measured the inhibition constants of each compound against HIV-1protease using the method described essentially by M. W. Pennington etal., Peptides 1990, Gimet, E. and D. Andrew, Eds., Escom; Leiden,Netherlands (1990).

Compounds of formula I were tested for their antiviral potency inseveral virological assays. In the first assay, the compounds were addedas a solution in dimethylsulfoxide (DMSO) to a test cell culture ofCCRM-CEM cells, a strain of CD4⁺ human T-cell lymphoma cells, previouslyacutely infected with HIV_(IIIb) using standard protocols (see Meek, T.D. et al., “Inhibition of HIV-1 protease in infected T-lymphocytes bysynthetic peptide analogues”, Nature, 343, p. 90 (1990). Preferredcompounds are those which are able to inhibit 90% of viral infectivityat a concentration of 1 μM or less. More preferred compounds are thosewhich are able to inhibit 90% of viral infectivity at a concentration of100 nM or less.

The effect of the compounds on inhibiting the replication of the viruswas measured by determining the HIV extracellular p24 antigenconcentration using a commercial enzyme immunoassay (obtained fromCoulter Corporation, Hialeah, Fla.).

Depending on the cell type and the desired readout, syncytia formation,reverse-transcriptase (RT) activity, or cytopathic effect as assayed bya dye uptake method may also be used as readouts of antiviral activity.See H. Mitsuya and S. Broder, “Inhibition of the in vitro infectivityand cytopathic effect of human T-lymphotropic virus typeIII/lymphoadenopathy-associated virus (HTLV-III/LAV) by2′,3′-dideoxynucleosides”, Proc. Natl. Acad. Sci. USA, vol. 83, pp.1911-1915 (1986). The effect of compounds of formula I on clinicalisolates of other HIV-1 strains was determined by obtaining low-passagedvirus from HIV-infected patients and assaying the effect of theinhibitors in preventing infection of the HIV virus in freshly preparedhuman peripheral blood mononuclear cells (PBMCs).

Insofar as compounds of formula I are able to inhibit the replication ofthe HIV virus in human T-cells and furthermore, may be delivered orallyto mammals, they are of evident clinical utility for the treatment ofHIV infection. These tests are predictive of the compounds ability toinhibit HIV protease in vivo.

EXAMPLE 1

A. Compound XI ((syn)-OH, D′=benzyl). 184 g of Brockman Super I gradeneutral alumina was slurried in sufficient diethyl ether to form athick, stirrable suspension and was treated with 7.48 mL of benzylamine.After stirring for 5 min, 7.28 g of(1S,2S)-1-(N-benzyoxycarbonyl)-amino-2-phenylethyl-oxirane was added andthe mixture stirred for 15 h. The mixture was treated with 15.28 g ofdi-tert-butylpyrocarbonate and 4.70 mL of diisopropylethylamine. Thismixture was stirred for 3.5 h, then treated with 600 mL of methanol,allowed to stand for 3.5 h, and filtered to yield a yellow oil, whichwas purified by silica gel chromatography using a gradient of 0.5 to1.5% methanol in methylene chloride to yield 3.88 g of the desiredproduct as a white solid. Further washing the filter cake with methanoland with 3% ammonium hydroxide in methanol yielded 2.2 g of4-benzylamino-2-N-benzyloxycarbonylamino-3-hydroxy-1-phenylbutane inseveral portions. Each of these portions was treated separately, as asolution in methylene chloride, with 1.1 molar equivalents each ofdi-tert butylpyrocarbonate and diisopropylethylamine, followed byaqueous workup with water, 10% aqueous KHSO₄, and brine, drying overMgSO₄, and concentration in vacuo. The combined products of thesereactions were purified by silica gel chromatography using a gradient of5% to 15% diethyl ether in methylene chloride. The resulting purefractions were collected and combined with the previously purifiedproduct to yield 5.49 g of a white solid. TLC: Rf=0.56, 5%methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XII ((syn)-OH, D′=benzyl). A solution of 5.49 g of theresultant compound of Example 1A in 40 mL of ethanol was hydrogenatedunder a slight positive pressure of hydrogen in the presence of 380 mgof 10% palladium on carbon for 16 h. After filtering and concentratingin vacuo, the desired product was obtained as 4.03 g of a white solid.TLC: Rf=0.21, 95:5:0.5 CH₂Cl₂/methanol/concentrated NH₄OH.

C. Compound XIII ((syn)-OH, A=benzyloxycarbonyl, D′=benzyl). A solutionof 3.02 g of the resultant compound of Example 1B in 150 mL of methylenechloride was treated with 4.35 g of N^(α)-Cbz-N^(δ)-trityl asparagine,1.16 g of hydroxybenzotriazole hydrate, and 1.64 g of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The mixturewas stirred for 16 h, then diluted with 3 volumes of diethyl ether andwashed sequentially with water, saturated NaHCO₃ solution, 10% KHSO₄solution, and brine. After drying over MgSO₄ and concentrating in vacuo,a yellow oil was obtained which was purified by chromatography on aFlorisil column using a gradient of 0% to 25% EtOAc in CH₂Cl₂ as eluantto yield 8.00 g of the title compound as a white foam. TLC: Rf=0.51, 5%methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

D. Compound XIV ((syn)-OH, A=H, D′=benzyl). A solution of 7.90 g of theresultant compound of Example 1C in 150 mL of ethanol was hydrogenatedunder a slight positive pressure of hydrogen in the presence of 550 mgof 10% palladium on carbon for 2.5 h, then ca. 50 mg more 10% palladiumon carbon was added, the mixture was then filtered and concentrated invacuo to give the desired product as 6.66 g of a white solid which wasused without subsequent purification. TLC: Rf=0.26, 95:5:0.5CH₂Cl₂/methanol/concentrated NH₄OH.

E. Compound XIV ((syn)-OH, A=quinoline-2-carbonyl, D=benzyl). Asuspension of 1.51 g of quinaldic acid and 6.17 g of the resultantcompound of Example 1D in 150 mL of acetonitrile was treated with 1.52mL of diisopropylethylamine and 3.58 g of BOP reagent. The mixture wasstirred for 14 h, then concentrated in vacuo. The gummy residue waspartitioned between ether and water, and the organic layer was washedsequentially with brine, saturated NaHCO₃ solution, water, 10% KHSO₄solution, and brine, then dried over MgSO₄ and concentrated in vacuo.Subsequent purification by silica gel chromatography using 0% to 8.5%solvent A in methylene chloride (where solvent A is defined as 90:10:1,methylene chloride/methanol/concentrated ammonium hydroxide) yielded5.79 g of the title compound as a white foam, along with ca. 600 mg ofslightly impure side fractions. TLC: Rf=0.41, 5% methanol/CH₂Cl₂;(¹H)-NMR (CDCl₃) consistent with structure.

F. Compound 1. A 58 mg portion of the resultant compound of Example 1Ewas treated with 1 mL of 90% aqueous TFA and allowed to stand for 17 h.The mixture was concentrated in vacuo and the residue taken up in 3 mLof CH₂Cl₂, treated with 100 μL of DIEA, and cooled to 0° C. To thissolution was added 26 μL of benzenesulfonyl chloride, and the mixturewas stirred for 18 h, warming slowly to ambient temperature. Afterconcentration of the mixture in vacuo, the residue was purified by thicklayer silica gel chromatography using 5% MeOH/CH₂Cl₂ as eluant followedby preparative reversed-phase C₁₈ HPLC using a linear gradient of 40% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 8.3 mg of the titlecompound. TLC: Rf=0.50, 5% MeOH/CH₂Cl₂. HPLC: Rt=17.8 min. NMR (DMSO-d₆)δ 2.62 (dd, 1H); 2.76 9d, 2H); 2.80 (dd, 1H); 3.11, (d, 2H); 3.34 (dd,1H); 4.59 (br s, 1H); 4.68 (br s, 1H); 3.97 (m, 1H); 4.20 (d, 1H), 4.35(d, 1H); 4.68 (dd, 1H); 6.39 (d, 1H); 6.74 (t, 1H); 6.81 (t, 2H); 6.93(d, 2H); 7.12-7.24 (m, 6H); 7.51 (t, 2H); 7.57 (t, 1H); 7.62 (dd, 1H);7.77 (t, 2H), 7.96 (d, 1H); 8.09 (d, 1H); 8.16 (d, 1H); 8.31 (d, 1H);8.53 (d, 1H).

EXAMPLE 2

Compound 2. A 150 mg portion of the resultant compound of Example 1E wasdissolved in 1 mL of 90% aqueous TFA and stirred at ambient temperatureovernight, then concentrated in vacuo. The crude TFA salt residue wasdissolved in 7 mL of dry methylene chloride and the pH of the solutionwas adjusted to pH 8 with 1N NaOH. 56 mg of a mixture of4-fluoro-3-acetamidobenzene sulfonylchloride and3-fluoro-4-acetamidobenzene sulfonylchloride (˜1:1) was added and themixture stirred vigorously for 3 hours after which an additional 25 mgwas added and the reaction allowed to continue for an additional 12hours. The reaction was then diluted with 50 mL of ethylene chloride andthe organic layer was washed sequentially with water and brine, driedover MgSO₄ and concentrated in vacuo. The crude residue was purifiedusing a silica gel flash chromatography column using a gradient of 3% to5% MeOH in methylene chloride as eluant to yield 60 mg of the titlecompounds. TLC: Rf=0.50, 10% MeOH/CH₂Cl₂; HPLC: Rt=13.93 min. NMR(CDCl₃): δ9.05 (s, 1H); 8.65 (d, 0.5H); 8.58 (t, 0.5H), 8.20 (dd, 0.5H),7.85 (d, 1H) 7.75 (m, 0.5H), 7.45-7.63 (m, 1.5H), 7.14-7.25 (m, 6H),6.78-6.95 (m, 5H), 6.70 (d, 1H), 6.41 (s, 0:5 H), 6.25 (s, 0.5H), 6.18(s, 0.5H), 6.10 (s, 0.5H), 4.88 (m, 0.5H), 4.81 (m, 0.5H), 4.37 (d, 1H),4.35 (m, 1H), 4.21 (d, 1H), 4.00 (m, 1H), 3.46 (m, 0.5H), 3.35 (m,0.5H), 3.27 (d, 0.5H), 3.16 (d, 0.5H), 3.14 (d, 1H), 2.45-2.75 (m, 5H);2.16, 2.20 (2 s, 3H total).

EXAMPLE 3

Compound 3. A 23 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h. Themixture was concentrated in vacuo and the residue taken upon in 2 mL ofCH₂Cl₂, treated with 6 μL of DIEA, and cooled to 0° C. To this solutionwas added 23 mg of 3,5-dimethylisoxazole-4-sulfonyl chloride, and themixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.1 mg of the titlecompound. TLC: Rf=0.55, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.5 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 4

Compound 4. A 33 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 15 h. Themixture was concentrated in vacuo and the residue taken up in 3 mL ofCH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To this solutionwas added 10 μL of 3-trifluoromethylbenzene sulfonyl chloride, and themixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.1 mg of the titlecompound. TLC: Rf=0.55, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.5 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 5

Compound 5. A 20 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 18 h. Themixture was concentrated in vacuo and the residue taken up in 1 mL ofCH₂Cl₂, treated with 10 μL of DIEA, and cooled to 0° C. To this solutionwas added 13 mg of 2-acetamido-4-methyl-5-thiazolesulfonyl chloride, andthe mixture was stirred for 17 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 0.40 mg of the titlecompound. TLC: Rf=0.5, 10% MeOH/CH₂Cl₂. HPLC: Rt=13.8 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 6

Compound 6. A 33 mg portion of the resultant compound of Example 1E wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 16 h. Themixture was concentrated in vacuo and the residue taken up in 2 mL ofCH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To this solutionwas added 11 mg of 5-(isoxazol-3-yl)thiophene-2-sulfonyl chloride, andthe mixture was stirred for 18 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 1.5 mg of the titlecompound. TLC: Rf=0.7, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.7 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 7

Compound 7. A 35.5 mg portion of the resultant compound of Example 1Ewas treated with 1 mL of 90% aqueous TFA and allowed to stand for 18 h.The mixture was concentrated in vacuo and the residue taken up in 3 mLof CH₂Cl₂, treated with 16 μL of DIEA, and cooled to 0° C. To thissolution was added 10 mg of 3-chlorosulfonylbenzoic acid, and themixture was stirred for 16 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% tTFA for elution to obtain 1.6 mg of the titlecompound. TLC: Rf=0.7, 10% MeOH/CH₂Cl₂. HPLC: Rt=13.6 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 8

Compound 8. 0.04 mmol of the resultant compound of Example 10A wasconverted to the free base by partitioning between EtOAc and sat.NaHCO₃. Treatment of the resulting compound with an excess of 1%HCl/MeOH and concentration in vacuo yielded the hydrochloride salt as awhite solid. This compound was suspended in CH₂Cl₂ and treated withsufficient DIEA to bring the pH to >10 (moist pH paper). The solutionwas treated with 7 molar equivalents of chlorotrimethylsilane andstirred for 15 h under nitrogen, then treated with 0.06 mmol of methanesulfonyl chloride and stirred for 1 h. The resulting mixture wasconcentrated to a small volume, applied directly to a thick layer silicagel plate and eluted with 7% MeOH/CH₂Cl₂. The primary UV-quenching bandwas isolated and further purified by preparative reversed-phase HPLC toyield the title compound as a white solid. TLC: Rf=0.65, 10%CH₃OH/CH₂Cl₂, HPLC: Rt=12.3 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLES 9 AND 192

A. Compound XIV ((syn, anti-OH, A=quinoline-2-carbonyl, D′=isobutyl). Asolution of 317 mg (0.425 mmol) of the resultant compounds of Example17B, diastereomer B and 0.11 mL (0.637 mmol) of diisopropylethyl aminein 7 mL of dichloromethane was treated with 139.1 mg (0.637 mmol) ofdi-tert-butyl dicarbonate. After 24 hours, the mixture was diluted withdichloromethane. The mixture was washed with water, 5% NaHCO₃, 0.5 NHCl, brine then dried over MgSO₄, filtered and concentrated in vacuo.The residue was purified by low pressure silica gel columnchromatography using a 20% ethyl acetate/dichloromethane as eluent toyield 81.2 mg of the fast moving hydroxyl diastereomer, 65.8 mg of theslower moving hydroxyl diastereomer, and 65.8 mg of the mixeddiastereomers. TLC: Rf=0.60, 0.67, 40% EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃)consistent with structure.

B. Compounds 9 and 192. A solution of 35.1 mg (0.041 mmol) of theresultant mixed diastereomers (˜1:1) of Example 9/192A in 0.8 mL ofdichloromethane was treated with 0.8 mL of trifluoroacetic acid. After 4hours, the mixture was concentrated in vacuo. TLC: Rf=0.11, 10%CH₃OH/CH₂Cl₂. To a solution of the resulting trifluoroacetic acid salt(entire yield) in 1 mL of dichloromethane was sequentialled added 0.3mLof saturated NaHCO₃, a small amount of solid NaHCO₃ and 11.8 mg (0.054mmol) of benzofurazan-4-sulphonyl chloride. After 3 hours, the mixturewas diluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 2.0 mg of compound 9 as a white solid: TLC: Rf=0.20, 5%CH₃OH/CH₂Cl₂; HPLC, RT 14.2 min. 2.7 mg of compound 192 was alsoobtained as a white solid, which was determined by NMR and HPLC to becontaminated with ˜25% of compound 9: TLC: Rf=0.20, 5% CH₃OH/CH₂Cl₂;HPLC, Rt=14.2 min. (¹H)-NMR consistent with structure.

EXAMPLE 10

A. Compound XV ((syn)-OH, A=quinoline-2-carbonyl, D′=benzyl; TFA salt).A 0° C. solution of 1.027 g portion of the resultant compound of Example1E in 5 mL of CH₂Cl₂ was treated with 5 mL of TFA and allowed to standfor 3 h. The mixture was concentrated in vacuo to yield 0.95 g of thetitle compound, which was used without subsequent purification.

B. Compound 10. A solution of 30.2 mg of the resultant compound ofExample 10A in 3 mL of CH₂Cl₂ was treated with 0.33 mL of DIEA and 31.1mg of m-benzenedisulfonyl chloride. The mixture was stirred for 2 h,then treated with 2 mL of concentrated aqueous ammonium hydroxide. Thebiphasic mixture was stirred for an additional 16 h, concentrated invacuo, and the residue partitioned between ethyl acetate and brine. Theorganic layer was dried over anhydrous MgSO₄ and concentrated in vacuo,and the residue was purified by preparative thick layer silica gelchromatography using 3% MeOH/CH₂Cl₂ as eluant to yield 4.5 mg of thetitle compound. TLC: Rf=0.5, 3% MeOH/CH₂Cl₂ as eluant to yield 4.5 mg ofthe title compound. TLC: Rf=0.5, 3% MeOH/CH₂Cl₂. HPLC: Rt=13.4 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 11

Compound 11. A solution of 57.9 mg of the resultant compound of Example10A in 5 mL of CH₂Cl₂ was treated with 30 μL of DIEA and 9.3 μL ofdimethylsulfamoyl chloride. The mixture was stirred for 12 h, thentreated with an additional 30 μL of DIEA and 9.3 μL of dimethylsulfamoylchloride and the reaction was allowed to proceed an additional 12 hours.The mixture was then diluted with CH₂Cl₂ and washed with saturatedNH₄Cl; the aqueous layer was washed with CH₂Cl₂, and the combinedorganic extracts were dried over MgSO₄. Filtration and concentrationprovided a residue which was chromatographed on a silica gel columnusing 2.5% MeOH/EtOAc as eluent, yielding a slightly impure productwhich was further purified by preparative HPLC using a linear gradientof 35% to 100% CH₃/CN/H₂O with 0.1% TFA for elution. HPLC: Rt=13.0minutes. NMR (CDCL₃): δ9.15 (d, 1H), 8.34 (d, 1H), 8.22 (d, 1H), 8.18(d, 1H), 7.90 (d, 1H), 7.80 (t, 1H), 7.65 (t, 1H), 7.16-738 (m, 5H),7.05 (d, 1H), 6.95 (t, 1H), 6.87 (t, 1H), 5.85 (br s, 1H), 5.62 (br s,1H), 4.87 (M, 1H), 4.46 (s, 2H), 4.08 (m, 1H), 3.66 (m, 1H), 3.30 (m,2H), 2.59-2.94 (m, 4H), 2.81 (s, 6H).

EXAMPLE 12

A. Compound XIV ((syn)-OH, A=quinoline-2-carbonyl, D′=benzyl;trifluoroacetate salt). To a solution of 1.027 g (1.164 mmol) of theresultant compound of Example 1E in CH₂Cl₂ (5 mL) at 0° to 5° C. wasadded trifluoromethanesulfonic acid (5 mL). After stirring for 3 h, thereaction mixture was concentrated in vacuo to provide 0.95 g of lightyellow, gummy product, containing one equivalent of triphenylmethanol,which was used without subsequent purification.

B. Compound 12. To a solution of 30.2 mg (0.038 mmol) of the resultantcompound of Example 12A in CH₂Cl₂ (3 mL) was added diisopropylethylamine(0.33 mL, 0.189 mmol), and 2-(pyrid-2-yl)-tyiophene-5-sulfonyl chloride13 mg, (0.249 mmol). After 14 h, the resulting mixture was diluted withethyl acetate, washed with saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified bypreparative reversed-phase chromatography using a 5% to 100%H₂O/acetonitrile gradient as eluant to yield the title product.

EXAMPLE 13

Compound 13. To a solution of 30 mg (0.038 mmol) of the resultantcompound of Example 12A in CH₂Cl₂ (3 mL) was added diisopropylethylamine(0.33 mL, 0.189 mmol), and 2-(3-phenylsulfonyl)thiophene sulfonylchloride (0.113 mmol). After stirring for 2 h, the reaction mixture wasmade biphasic by addition of 30% ammonium hydroxide solution (2 mL).After stirring for an additional 16 h, the resultant mixture wasconcentrated in vacuo, reconstituted in ethyl acetate, washed withsaturated brine, dried over magnesium sulfate, filtered, andre-concentrated in vacuo. Purification by thin layer preparativechromatography yielded the desired compound.

EXAMPLE 14

Compound 14. The resulting compound of Example 17B, diastereomer B (170mg) was treated with 1 mL of 90% aqueous TFA and allowed to stand for 12h. The mixture was concentrated in vacuo and the residue is taken up in5 mL of dry CH₂Cl₂. To this solution, 3 mL of saturated aqueous sodiumbicarbonate and 50 mg of 4-fluorobenzenesulfonyl chloride was added andthe mixture stirred for 3 h. The resulting mixture was diluted withCH₂Cl₂ and washed with water, dried over magnesium sulfate and filtered.After concentration of the mixture in vacuo, a portion of the residuewas purified by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution to obtain3.0 mg of the title compound. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=14.78 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 15

Compound 15. A sample of a mixture of4-fluoro-3-acetamidobenzenesulfonyl chloride and3-fluoro-4-acetamidobenzenesulfonyl chloride (approx. 1:1; obtained fromMaybridge Chemicals) was resolved into its respective regioisomers bysilica gel chromatography using 10% isopropyl alcohol/hexane as eluent.A solution of 4-acetamido-3-fluorobenzenesulfonyl chloride (30 mg) andthe resulting compound of Example 17B, diastereomer B (80 mg) in 10 mLof CH₂Cl₂ was reacted in the same manner as described for Example 14.After workup and purification of a portion of the product by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluent, 1.2 mg of the title compound was obtained as awhite solid. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=12.91 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 16

Compound 16. 80 mg of the resulting compound of Example 17B,diastereomer B, was reacted with 45 mg of3-acetamido-4-fluorobenzenesulfonyl chloride in the same manner asdescribed for Example 14. After workup and purification of a portion ofthe product by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 1.4 mg of thetitle compound was obtained. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=12.91 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 17

A. (2S)-2-((1S,2R syn,anti)-3-(2-methylpropyl)amino-1-benzyl-2-hydroxypropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-tritylsuccinamide. A solution of 683.1 mg (0.96 mmol) of the resultantcompounds of Example 191D and 1.9 mL (19.2 mmol) of isobutylamine in 10mL of acetonitrile in a sealed tube was heated at 90-100° C. for 24hours. After cooling to room temperature, the mixture was concentratedin vacuo. The residue was taken up in dichloromethane and washed withwater, brine, then dried over MgSO₄, filtered and concentrated in vacuoto yield 783.8 mg of the mixed diastereomeric products. TLC: Rf=0.11,10% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XIII, ((syn, anti)-OH, P=quinoline-2-carbonyl, D′=isobutyl).A solution of 583.8 mg of the resultant compounds of Example 17A and 0.2mL of diisopropylethylamine in 10 mL of dichloromethane was treated with256 mg of di-tert-butyl dicarbonate. After 24 hours, the mixture wasdiluted with dichloromethane. The mixture was washed with water, 5%NaHCO3, 0.5 N HCl, brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by low pressure silicagel column chromatography using a 20% ethyl acetate/dichloromethane aseluent to yield 154.6 mg of the fast moving diastereomer A, lateridentified as having the anti configuration at the hydroxyl center; 98.8mg of the slower moving diastereomer B, having the syn configuration atthe hydroxyl center, and 204.6 mg of the mixed diastereomers A and B.TLC: Rf=0.60, 0.67, 40% EtOAc/CH₂Cl₂.

C. Compound 17. A solution of 64.6 mg of the resultant compounds ofExample 17B, diastereomer B, in 1.5 mL of dichloromethane was treatedwith 1.5 mL of trifluoroacetic acid. After 4 hours, the mixture wasconcentrated in vacuo to yield the amine trifluoroacetate salt. TLC:Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solution of 17.8 mg of the resultanttrifluoroacetate salt in 1 mL of dichloromethane was sequentially added0.3 mL of saturated NaHCO₃, a small amount of solid NaHCO₃ and 10.7 mgof 4-acetamido-benzenesulphonyl chloride. After 3 hours, the mixture wasdiluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 14.4 mg of the title compound as a white solid; TLC: Rf=0.54, 10%CH₃OH/CH₂Cl₂; HPLC, Rt=13.58 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 18

Compound 18. To a solution of 20.8 mg (0.041 mmol) of the crudetrifluoroacetate salt obtained as from Example 17B, diastereomer B, in 1mL of dichloromethane was sequentially added 0.3 mL of saturated NaHCO₃,a small amount of solid NaHCO₃ and 13.6 mg (0.054 mmol) of2-acetamido-4-methyl-5-thiazolesulphonyl chloride. After 3 hours, themixture was diluted with dichloromethane. The two layers were separatedand the aqueous layer was extracted once with dichloromethane. Thecombined organic layer was washed with brine then dried over MgSO₄,filtered and concentrated in vacuo. The residue was purified bypreparative HPLC to yield 4.8 mg of the title compound as a white solid;TLC: Rf=0.50, 10% CH₃OH/CH₂Cl₂; HPLC: Rt=13.35 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 19

A. Sodium 3-acetamidobenzenesulfonate. A solution of 118.6 mg (0.55mmol) of 3-acetamidobenzenesulfonic acid in 0.5 mL of water was treatedwith 0.55 mL (0.55 mmol) of 1.0 N NaOH at 0° C. After stirring at roomtemperature for 4 hours, the mixture was concentrated to dryness andused without subsequent purification.

B. 3-Acetamidobenzenesulfonyl chloride. The crude mixture from Example19A was cooled to 0° C. and 0.29 g (1.38 mmol) of phosphoruspentachloride was added. The mixture of solid was stirred for 3 hoursthen 5 mL dichloromethane was added. After 24 hours, the slurry wasfiltered and concentrated in vacuo to yield 81.4 mg of solid productwhich was used without subsequent purification. TLC: Rf=0.50, 40%EtOAC/CH₂Cl₂.

C. Compound 19. A solution of 82.7 mg (0.098 mmol) of diastereomer B,obtained in Example 17B, in 2 mL of dichloromethane was treated with 2mL of trifluoroacetic acid. After 4 hours, the mixture was concentratedin vacuo to yield the amine trifluoroacetate salt which was used withoutfurther purification; TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. A solution of thissalt (entire yield) in 2 mL of dichloromethane was treated sequentiallywith 0.5 mL of saturated NaHCHO₃, small amount of solid NaHCO₃ and asolution of 81.4 mg (0.046 mmol) of the resultant compound of Example19B. After 3 hours, the mixture was diluted with dichloromethane. Thetwo layers were separated and the aqueous layer was extracted once withdichloromethane. The combined organic layer was washed with brine thendried over MgSO₄, filtered and concentrated in vacuo. The residue waspurified by preparative HPLC to yield 24.7 mg of the title compound as awhite solid; TLC: Rf=0.42, 10% CH₃OH/CH₂Cl₂; HPLC: Rt=13.8 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 20

Compound 20. A solution of 209.0 mg (0.24 mmol) of the resultantcompound of Example 17B, diastereomer B, in 5 mL of dichloromethane wastreated with 5 mL of trifluoroacetic acid. After 4 hours, the mixturewas concentrated in vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solutionof this residue in 2 mL of dichloromethane was sequentially added 0.5 mLof saturated NaHCO₃, a small amount of solid NaHCO₃ and 70.2 mg (0.32mmol) of benzofurazan-4-sulphonyl chloride. After 3 hours, the mixturewas diluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 108.0 mg of the title compound as a white solid; TLC: Rf=0.60, 10%CH₃OH/CH₂Cl₂; HPLC: Rt=14.95 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 21

Compound 21. The resulting compound of Example 17B, diastereomer B, (228mg, 0.27 mmol) was dissolved in 1:1 CH₂Cl₂/TFA (10 mL), and the reactionmixture stirred for 3.5 hours, then concentrated to dryness to affordthe product trifluoroacetate salt as a yellow solid which was used inthe next reaction without purification. To a solution of this residue(34.7 mg, 0.05 mmol) in CH₂Cl₂ (3 mL) was added Heunig's base (41 μl,0.24 mmol) and dimethylsulfamoyl chloride (11 μl, 0.09 mmol), and thereaction was stirred for 17 hours at room temperature. The reactionmixture was then diluted with CH₂Cl₂ and washed with saturated NH₄Cl,and the organic layer was dried over MgSO₄. Filtration and concentrationprovided a residue which was chromatographed on a silica gel columnusing 8% CH₃OH/CH₂Cl₂ as eluent, yielding the desired compound which wasfurther subject to purification by preparative HPLC. HPLC: Rt=13.8minutes. TLC: Rf=0.40, 8% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 22

A. N^(α)-isocyano-L-valine methyl ester. To the HCl salt of valinemethyl ester (2.08 g, 12.40 mmol) in toluene (20 mL) was added a 20%solution of phosgene in toluene (32 mL, 62.00 mmol), and the solutionwas heated at reflux for 12 hours. The reaction was then cooled to roomtemperature and concentrated in vacuo to give a pale yellow liquid whichwas used in the subsequent reaction without purification. TLC: Rf=0.88,50% Hexane/EtOAc; (¹H)-NMR (CDCl₃) consistent with structure.

B. N^(α)-(2-pyridylmethyl)-oxycarbonyl-L-valine methyl ester. A mixtureof 2-pyridylcarbinol (941 μl, 9.75 mmol) and the resulting compound ofExample 22A (1.28 g, 8.12 mmol) were allowed to stir in CH₂Cl₂ (7 mL)for 12 hours, then the reaction was concentrated and the residuechromatographed with 50% hexane/EtOAc to afford 2.03 grams of the titlecompound as a colorless oil. TLC: Rf=0.26, 50% Hexane/EtOAc; (¹H)-NMR(CDCl₃) consistent with structure.

C. N^(α)-(2-pyridylmethyl)-oxycarbonyl-L-valine. A solution of theresulting compound of Example 22B (634 mg, 2.38 mmol) in a 1/1 mixtureof 1N HCl/THF (6 mL) containing 12 N HCl (0.5 mL) was allowed to stir atroom temperature over 15 hours, but much starting material was stillpresent by TLC. Hence, more 12 N HCL was added (1 mL), and the reactionstirred an additional 48 hours. The reaction was then concentrated todryness and diluted with CH₂Cl₂, yielding the desired carboxylic acid asan insoluble resin which was washed with additional CH₂Cl₂, providing22C which contained minor quantities of 22B. This material was used inthe subsequent reaction without further purification. TLC: Rf=0.11, 8%CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

D. Compound XXX (P=(2-pyridylmethyl)-oxycarbonyl, R³=isopropyl,R^(3′)═H, D′=isobutyl, P′=tert-butoxycarbonyl). To the resultingcompound of Example 21B (277 mg, 0.82 mmol) in CH₂Cl₂ (5 mL) was added1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (210 mg,1.10 mmol), the acid 22C (402 mg, 1.10 mmol), and 1-hydroxybenzotriazolehydrate (148 mg, 1.10 mmol). The reaction proceeded for 12 hours at roomtemperature, then was diluted with CH₂Cl₂ and washed successively withsaturated NH₄Cl and NaHCO₃, and the organic layer was dried over MgSO₄.Filtration and concentration provided a residue which waschromatographed on a silica gel column using 17% THF/CH₂Cl₂ as eluent,yielding 396 mg of product. TLC: Rf=0.26, 17% THF/CH₂Cl₂; (¹H)-NMR(CDCl₃) consistent with structure.

E. Compound 22. The resulting compound of Example 22D (396 mg, 0.69mmol) was dissolved in 90% aqueous TFA (11 mL), and the reaction mixturestirred for 3 hours at room temperature, then was concentrated todryness. To a solution of this residue (231 mg. 0.33 mmol) in CH₂Cl₂ (5mL) was added excess solid NaHCO₃ (approx. 1 gram) and saturated aqueousNaHCO₃ (20 μl), followed by N-acetylsulfanilyl chloride (116 mg, 0.50mmol), and the reaction proceeded for 12 hours at room temperature. Thereaction mixture was then diluted with CH₂Cl₂ and washed with saturatedNaHCO₃, and the organic layer was dried over MgSO₄. Filtration andconcentration provided a residue which was chromatographed on a silicagel column using 8% CH₃OH/CH₂Cl₂ as eluent, yielding the desiredcompound which was further subject to purification by preparative HPLC(76.1 mg of 3 was obtained). HPLC: Rt=12.1 minutes. TLC: Rf=0.46, 8%CH₃OH/CH₂Cl₂; NMR (CDCl₃) 8.76 (d, 1H), 8.40 (br s, 1H), 8.26 (t, 1H),7.72 (d, 2H), 7.67 (d, 2H), 7.58 (d, 2H), 7.37 (d, 1H), 7.25 (m, 4H),7.16 (br d, 1H), 6.47 (d, 1H), 5.65 (d, 1H), 5.26 (d, 1H), 4.32 (m, 1H),3.91 (t, 1H), 3.83 (m, 1H), 3.23 (d, 1H), 3.05 (m, 2H), 2.68-3.10 (m,3H), 2.22 (m, 3H), 2.0 (m, 1H), 1.82 (m, 1H), 0.85 (d, 3H), 0.80 (d,3H), 0.71 (d, 3H), 0.65 (d, 3H).

EXAMPLE 23

Compound 23. Prepared by the same route as described for Example 22,except 4-pyridylcarbinol was utilized for reaction with the product ofExample 22A. HPLC: Rt=12.0 minutes. TLC: Rf=0.50 (8% CH₃OH/CH₂Cl₂);(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 24

Compound 24. A solution of the resulting compound of the trifluoroaceticacid deprotection of Example 22D (as described in Example 22E; 215 mg,0.31 mmol) in CH₂Cl₂ at room temperature was treated withdiisopropylethylamine (214 μl, 1.23 mmol) and dimethylsulfamoyl chloride(40 μl, 0.37 mmol) in CH₂Cl₂ at room temperature in CH₂Cl₂ at roomtemperature for 12 hours. The reaction mixture was concentrated andchromatographed on a silica gel column with 5% CH₃OH/CH₂Cl₂ as eluent,yielding the desired compound which was further subject to purificationby preparative HPLC (9.5 mg obtained). HPLC: Rt=14.4 minutes. TLC:Rf=0.88, 11% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 25

Compound 25. This compound was prepared by the route described forExample 22, except that 3-pyridylcarbinol was utilized for reaction withthe compound produced in Example 22A, and in the reaction correspondingto 22E, the trifluoracetate-deprotected material was reacted withbenzofurazan-4-sulphonyl chloride. HPLC: Rt=9.4 minutes. TLC: Rf=0.10,11% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 26

Compound 26. A solution of the resulting compound from thetrifluoroacetic acid deprotection of Example 22D (as described inExample 22E; 27 mg, 0.14 mmol) in CH₂Cl₂ was treated with excess solidNaHCO₃ (approx. 1 gram) and saturated aqueous NaHCO₃ (7 μl), thenstirred vigorously at room temperature for 3 hours. The reaction mixturewas decanted from the solids, concentrated, then the residue waspurified directly by preparation HPLC (3.0 mg of white solid obtained).HPLC: Rt=14.7 minutes; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 27

Compound 27. A solution of 33 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 20 mg of N,N-diisopropylethylamine and 9.3mg of allyl chloroformate. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 24 mg of the titlecompound as a white solid. TLC: Rf=0.53, 5:10:85 NH₄H/CH₃OH/CH₂Cl₂.HPLC: Rt=14.53 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 28

Compound 28. A solution of 47.5 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 28.7 mg of N,N-diisopropylethylamine and15.2 mg of isobutyl chloroformate. The mixture was stirred 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 45 mg of the titlecompound as a white solid. TLC: Rf=0.60, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=15.58 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 29

Compound 29. A solution of 35.6 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 21.5 mg of N,N-diisopropylethylamine and0.083 mL of 1.0 M isopropyl chloroformate. The mixture was stirred 3 hand then concentrated in vacuo. The residue was taken up in ethylacetate and washed with 0.5 N HCl and saturated NaCl then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethyl ether to yield 33.2 mg of the titlecompound as a white solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=14.81 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 30

A. (2-Pyrrolidinonyl-hydroxyethyl-N-hydroxysuccinimidyl carbonate. Asolution of 572 mg of 1-(2-hydroxyethyl)-2-pyrrolidinone and 1.70 g ofN,N′-disuccinimidyl carbonate in acetonitrile was treated, at ambienttemperature under an atmosphere of nitrogen, with 1717 mg ofN,N-diisopropylethylamine. The mixture was stirred for 14 h andconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with saturated NaHCO₃, saturated NaCl, then dried over MgSO₄,filtered, and concentrated in vacuo to yield 200 mg of a white solid.TLC: Rf=0.56, 10% isopropanol in CH₂Cl₂; (¹H)-NMR (CDCl₃) consistentwith structure.

B. Compound 30. A solution of 68 mg of the resultant compound of Example30A in CH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, to a solution of 32 mg of the resultant compound of Example40A and 39 mg N,N-diisopropylethylamine in CH₂Cl₂. The mixture wasstirred for 4 h, diluted with CH₂Cl₂, washed with saturated NaHCO₃ andsaturated NaCl, then dried over MgSO₄, filtered, and concentrated invacuo. The residue was subjected to preparative thin layer silica gelchromatography using a 2:1 mixture of 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethylether to yield 45 mg of residue. About 20 mg of this residue waspurified by preparative HPLC to yield 13.5 mg of the title compound as awhite solid. TLC: Rf=0.47, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=12.79mid; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 31

Compound 31. A solution of 39.7 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 24 mg of N,N-diisopropylethylamine and 14.5mg of phenyl chloroformate. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂:diethyl ether to yield 39.7 mg of the titlecompound. TLC: Rf=0.53, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=15.22 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 32

Compound 32. A solution of 391 mg of the resultant compound of Example39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 271 mg of4-fluorobenzenesulfonyl chloride and 117 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaCl then dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by low pressure silica gel chromatography using 5%diethyl ether in CH₂Cl₂ as eluent to yield 420 mg of the title compoundas a white solid. TLC: Rf=0.20, 5% diethyl ether in CH₂Cl₂. HPLC:Rt=17.41 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 33

Compound 33. A solution of 30 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 18.1 mg of N,N-diisopropylethylamine and9.3 mg of benzyl isocyanate. The mixture was stirred 14 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a mixture of5:10:85 NH₄OH/CH₃OH/CH₂Cl₂ to yield 30.2 mg of the title compound as awhite solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂. HPLC: Rt=14.36min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 34

Compound 34. A solution of 55 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 33.3 mg of N,N-diisopropylethylamine and17.8 mg of 2-methoxyethyl chloroformate. The mixture was stirred for 3 hand then concentrated in vacuo. The residue was taken up in ethylacetate and washed with 0.5 N HCl and saturated NaCl then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 48.1 mg of the titlecompound as a white solid. TLC: Rf=0.56, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂.HPLC: Rt=13.43 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 35

A. Compound XXI (D′=isobutyl, A′=4-fluorophenyl, hydrochloride salt). Asolution of 398 mg of the resultant compound of Example 32 in ethylacetate was treated at −20° C. with HCl gas. The HCl was bubbled throughthe mixture for 20 min over which time the temperature was allowed towarm to 20° C. Nitrogen was then bubbled through the mixture for 15 minand solvent removed in vacuo to yield 347 mg of the title compound as awhite solid. TLC: Rf=0.82, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃)consistent with structure.

B. Compound 35. A solution of 111 mg of the resultant compound ofExample 35A in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, to a solution of 118 mg of the resultantcompound of Example 48A and 133 mg N,N-diisopropylethylamine in CH₂Cl₂.The mixture was stirred for 14 h, diluted with CH₂Cl₂, washed withsaturated NaHCO₃ and saturated NaCl, then dried over MgSO₄, filtered,and concentrated in vacuo. The residue was subjected to preparative thinlayer silica gel chromatography using 5% CH₃OH in CH₂Cl₂ to yield 98.8mg of the title compound as a white solid. TLC: Rf=0.48, 5% CH₃OH inCH₂Cl₂. HPLC: Rt=15.18 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 36

Compound 36. A solution of 48 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 29.0 mg of N,N-diisopropylethylamine and15.1 mg of 3-butenyl chloroformate. The mixture was stirred for 3 h andthen concentrated in vacuo. The residue was taken up in ethyl acetateand washed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using a 2:1 mixture of(5:10:85 NH₄OH/CH₃OH/CH₂Cl₂):diethyl ether to yield 43.8 mg of the titlecompound as a white solid. TLC: Rf=0.83, 5:10:85 NH₄OH/CH₃OH/CH₂Cl₂;Rf=0.24, 5% diethyl ether in CH₂Cl₂. HPLC: Rt=14.76 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 37

Compound 37. A solution of 99 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 83.2 mg of3,4-dichlorobenzenesulfonyl chloride and 29 mg of sodium bicarbonate.The mixture was stirred for 14 h, diluted with CH₂Cl₂, washed withsaturated NaCl then dried over MgSO₄, filtered, and concentrated invacuo. The residue was subjected to operative thin layer silica gelchromatography using 5% CH₃OH in CH₂Cl₂ to yield 107 mg of the titlecompound as a white solid. TLC: Rf=0.35 (5% CH₃OH in CH₂Cl₂). HPLC:Rt=17.27 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 38

Compound 38. To a solution of 32 mg of the resultant compound of Example35A in CH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, 14 mg of benzyl chloroformate and 21 mgN,N-diisopropylethylamine. The mixture was stirred for 4 h, diluted withCH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using 10% diethyl etherin CH₂Cl₂ as eluent to yield 33 mg of product. TLC: Rf=0.62, 10% diethylether in CH₂Cl₂. HPLC: Rt=17.27 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 39

A. Compound XXI (D′=isobutyl, P=tert-butoxy carbonyl, P′═H). A solutionof 4.1 g of epoxide XX (P=Boc) in 30 mL of ethanol was treated with 22.4mL of isobutylamine and heated under reflux for 1 h. The mixture wasconcentrated to yield the title compound as a white solid which was usedwithout subsequent purification. NMR (CDCl₃): δ0.91 (d, 3H); 0.93 (d,3H); 1.37 (s, 9H); 1.68 (br s, 2H); 2.40 (d, 2H); 2.68 (d, 2H); 2.87(dd, 1H); 2.99 (dd, 1H); 3.46 (dd, 1H); 3.75 (br s, 1H); 3.80 (br s,1H); 4.69 (d, 1H); 7.19-7.32 (m, 4H).

B. Compound 39. To a solution of 514.1 mg of the resultant compound ofExample 39A in dichloromethane (10 mL) was added aqueous sodiumbicarbonate (5 mL) and N-acetylsulfanilyl chloride (428.4 mg). After 14h, the resulting mixture was diluted with ethyl acetate, washed withsodium bicarbonate, saturated brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 20% ethyl acetate indichloromethane eluent to yield 714.4 mg of the title product. TLC:Rf=0.63, 60% ethyl acetate/dichloromethane, HPLC: Rt=15.3 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 40

A. Compound XXII (D′=isobutyl, P═H, E=4-acetamidophenyl), hydrochloridesalt. To a solution of 691.4 mg (1.296 mmol) of the resultant compoundof Example 39B in ethyl acetate (20 mL) at −20° C. was bubbled anhydrousHCl gas for 10 min. The ice bath was removed and after an additional 15min., the reaction mixture was sparged with nitrogen then concentratedin vacuo to provide 610 mg of title product which was used withoutsubsequent purification.

B. Compound 40. A solution of 41.5 mg of the resultant crude compound ofExample 40A in 5 mL of dichloromethane was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 18.1 mg ofL-dihydroorotic acid, 0.031 mL (0.176 mmol) diisopropylethylamine, 15.5mg (0.115 mmol) of 1-hydroxybenzotriazole hydrate, 22 mg (0.115 mmol)EDC. After 1 h, the slurry was treated with 1 mL of dimethylformamide.The mixture was stirred for 16 h and then concentrated in vacuo. Theresidue was taken up in ethyl acetate and washed with water andsaturated brine, dried over magnesium sulfate, filtered and concentratedin vacuo. The residue was purified by thin layer preparativechromatography using (1/2/17 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane) eluent to provide 34.2 mg of the titleproduct. TLC: Rf=0.33, 1/2/17 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane). HPLC: Rt=11.3 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 41

Compound 4-1. To a solution of 42.8 mg of the resultant compound ofExample 40A in 5 mL dichloromethane was added sequentially, at ambienttemperature under an atmosphere of nitrogen, 17.2 mg of N-tert-butylglyoxalic acid, 0.032 mL diisopropylethylamine, 16 mg of1-hydroxybenzotriazole hydrate, 22.6 mg EDC. The mixture was stirred for16 h and then concentrated in vacuo. The residue was taken up in ethylacetate and washed with water, 0.5 N hydrochloric acid, washed withsodium bicarbonate, saturated brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by thinlayer preparative chromatography using 40% ethyl acetate/dichloromethaneeluent to provide 14.9 mg of the title product. TLC: Rf=0.47, 40% ethylacetate/dichloromethane, HPLC: Rt=15.2 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 42

Compound 42. To a solution of 43.5 mg of the resultant crude compound ofExample 40A in 5 mL dichloromethane was added sequentially at ambienttemperature, under an atmosphere of nitrogen, 13.0 mg of succinamicacid, 0.024 mL diisopropylethylamine, 15.0 mg of 1-Hydroxybenzotriazolehydrate, and 21.3 mg EDC. The mixture was stirred for 16 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with sodium bicarbonate, saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified bythin layer preparative chromatography using (1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane) eluent to provide 35.3 mg of the titleproduct. TLC: Rf=0.25, 1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane, HPLC: Rt=11.6 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 43

Compound 43. To a solution of 42.8 mg of the resultant compound ofExample 40A in 5 mL dichloromethane was added sequentially, at ambienttemperature under an atmosphere of nitrogen, with 14.1 mg ofL-pyroglutamic acid, 0.024 mL diisopropylethylamine, 14.8 mg of1-hydroxybenzotriazole hydrate, 20.9 mg EDC. The mixture was stirred for16 h and then concentrated in vacuo. The residue was taken up in ethylacetate and washed with water, 0.5 N hydrochloric acid, washed withsodium bicarbonate, saturated brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by thinlayer preparative chromatography using (1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane) eluent to provide 29.9 mg of the titleproduct. TLC: Rf=0.33, 1/2/11 v/v/v/30% ammoniumhydroxide/methanol/dichlomethane, HPLC: Rt=11.7 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 44

A. 3-Pyridylmethyl-N-hydroxysuccinimidyl carbonate. To a solution of181.0 mg of 3 pyidinecarbinol in 5 mL acetonitrile was addedsequentially at ambient temperature under an atmosphere of nitrogen,with 0.72 mL diisopropylethylamine and 354.1 mg of N,N′-disuccinimidylcarbonate. After 4 h, the resultant mixture was concentrated in vacuo toprovide a yellow solid which was used without subsequent purification.

B. Compound 44. To a solution of 58.1 mg of the resultant crude compoundof Example 40A in 3 mL of dichloromethane was added sequentially, atambient temperature under an atmosphere of nitrogen, 0.075 mLdiisopropylethylamine and 46.3 mg of the resultant compound of Example20A. The mixture was stirred for 16 h and then concentrated in vacuo.The residue was taken up in diethyl ether and extracted into 3×25 mL of0.5N HCl. The combined aqueous extracts were adjusted to pH 8 with solidsodium bicarbonate and extracted into 3×25 mL ethyl acetate. Thecombined organic extracts were washed with saturated brine, dried overmagnesium sulfate, filtered, and concentrated in vacuo. The residue waspurified by thin layer preparative chromatography using (1/2/17/20v/v/v/30% ammonium hydroxide/methanol/dichlomethane/diethyl ether)eluent to provide 10.3 mg of the title product. TLC: Rf=0.4, 1/2/17/20v/v/v/30% ammonium hydroxide/methanol/dichlomethane/diethyl ether, HPLC:Rt=11.8 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 45

Compound 45. To a solution of 28.3 mg of the resultant compound ofExample 39A in 4 mL of dichloromethane was added 1 mL saturated aqueoussodium bicarbonate solution, 9.2 mg sodium bicarbonate, and 0.013 mL ofbenzenesulfonyl chloride. After 14 h, the resulting mixture was dilutedwith ethyl acetate, washed with saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified bythin layer preparative chromatography using 10% diethylether/dichloromethane eluent to provide 19.3 mg of the title product.TLC: Rf=0.84, 25% diethyl ether/dichlormethane, HPLC: Rt=17.2 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 46

Compound 46. To a solution of 47.0 mg (0.140 mmol) of the resultantcompound of Example 39A in 4 mL of dichloromethane was added 1 mLsaturated aqueous sodium bicarbonate solution, 17.6 mg of solid sodiumbicarbonate, and 41.4 mg of 2,4 dimethylthiazole-5-sulfonyl chloride.After 14 h, the resulting mixture was diluted with ethyl acetate, washedwith saturated brine, dried over magnesium sulfate, filtered and isconcentrated in vacuo. The residue was purified by thin layerpreparative chromatography using 25% ethyl acetate/dichloromethaneeluent to provide 34.6 mg of the title product. TLC: Rf=0.44, 25%diethyl ether/dichloromethane, HPLC: Rt=16.4 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 47

Compound 47. To a solution of 50.7 mg of the resultant compound ofExample 39A in 4 mL of dichloromethane was added 1 mL saturated aqueoussodium bicarbonate solution, 15.2 mg of solid sodium bicarbonate, and2-fluorobenzenesulfonyl chloride 35.2 mg. After 14 h, the resultingmixture was diluted with ethyl acetate, washed with saturated brine,dried over magnesium sulfate, filtered and concentrated in vacuo. Theresidue was purified by thin layer preparative chromatography using 10%diethyl ether/dichloromethane eluent to provide 40.5 mg of the titleproduct. TLC: Rf=0.44, 25% diethyl ether/dichloromethane, HPLC: Rt=17.2min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 48

A. N-succinimidyl-(S)-3-tetrahydrofuryl carbonate. To a solution of 12.5mL of 1.93 M phosgene in toluene at 0-5° C. was added 1.3 g of(S)-(+)-3-hydroxy-tetrahydrofuran. After stirring for 2 h, the reactionmixture was sparged with nitrogen and then concentrated to dryness invacuo to provide 1.486 g of crude chloroformate. This material was takenup in 10 mL of acetonitrile and treated sequentially at ambienttemperature under an atmosphere of nitrogen with 1.17 g ofN-hydroxysuccinimide and 1.41 mL of triethylamine. After stirring for 14h, the reaction mixture was concentrated in vacuo to provide 3.44 g ofthe title product as a white solid.

B. Compound 48. To a solution of 87.2 mg of the resultant compound ofExample 40A in 5 mL of dichloromethane was added sequentially, atambient temperature under an atmosphere of nitrogen, 0.113 mLdiisopropylethylamine and 68 mg of the resultant compound of Example48A. The mixture was stirred for 16 h and then concentrated in vacuo.The residue was taken up in ethyl acetate and washed with water, 0.5 NHCl, saturated sodium bicarbonate, saturated brine, dried over magnesiumsulfate, filtered, and concentrated in vacuo. The residue was purifiedby silica gel chromatography using (3/6/20/65 v/v/v/v/30% ammoniumhydroxide/methanol/diethyl ether/dichlomethane) eluent followed bycrystallization from a mixture of dichloromethane, diethyl ether, andhexanes to provide 58 mg of the title product. TLC: Rf=0.17, 75% ethylacetate/dichloromethane, HPLC: Rt=13.1 min.; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 49

Compound 49. Following the procedure described in Example 83, a solutionof the resultant compound of Example 39A in CH₂Cl₂ is reacted with2,4-difluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄ filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 50

Compound 50. A solution of 30 mg of the resulting compound of Example 58and 9 μL of dimethysulfamoyl chloride in 10 mL of CH₂Cl₂ was reacted inthe same manner as described for Example 14. After workup andpurification by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 6.5 mg of thetitle compound was obtained. TLC: Rf=0.2, 3% CH₃OH in CH₂Cl₂. HPLC:Rt=15.96 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 51

A. Compound XXI (A=tert-butoxycarbonyl, D′=isobutyl,A′=benzyloxycarbonyl). To a solution of the resultant compound ofExample 39A (2.5 g, 7.43 mmol) in CH₂Cl₂ (50 mL) was added triethylamine(2.1 mL, 14.9 mmol) followed by addition of benzyl chloroformate (1.2mL, 8.1 mmol). The mixture was allowed to stir at ambient temperaturefor 6 h. The solution was diluted with 1 L of CH₂Cl₂ and washed withwater. The organics were dried over anhydrous MgSO₄, concentrated underreduced pressure, then purified via silica gel chromatography. Gradientsolvent system: CH₂Cl₂ followed by 3:97 methanol/CH₂Cl₂. The titlecompound (2.97 g, was obtained as a colorless oil. TLC: Rf=0.14, 3:97methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXI (A=H, D′=isobutyl, A′=benzyloxycarbonyl, hydrochloridesalt). To a solution of 1.5 g (3.187 mmol) of the resultant compound ofExample 51A in ethyl acetate (25 mL) at −20° C. was bubbled anhydrousHCl gas for 10 min. The ice bath was removed and after an additional 15min. the reaction mixture was sparged with nitrogen, then concentratedin vacuo to provide 1.29 g of title product as a white solid which wasused directly for ensuing reaction. TLC: Rf=0.14, 10% methanol/CH₂Cl₂.

C. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl, D′=isobutyl,A′=benzyloxycarbonyl). To a solution of 1.077 g of the resultant crudecompound of Example 51B (2.647 mmol) in acetonitrile (10 mL) was addedsequentially at ambient temperature under an atmosphere of nitrogen,1.61 mL (9.263 mmol) of diisopropylethylamine and 910 mg (3.97 mmol) ofthe resultant compound of Example 48A. After stirring for 3 h, anadditional 223 mg (0.973 mmol) of the resultant compound of Example 48Awas added. The mixture was stirred for 16 h and then concentrated invacuo. The residue was taken up in ethyl acetate and washed with water,0.5 N HCl, saturated sodium bicarbonate, saturated brine, dried overmagnesium sulfate, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel column chromatography using agradient 10% to 25% ethyl acetate in CH₂Cl₂ eluent to yield 1.025 g ofthe title product as a white solid. TLC: Rf=0.10, 10% ethylacetate/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

D. Compound XXI (A=(S)-3-tetrahydrofuryloxycarbonyl, D=isobutyl, A′=H).A solution of 872 mg (1.799 mmol) of the resultant compounds of Example51C in (10 mL) of ethyl alcohol was added, at ambient temperature undera nitrogen atmosphere, to a slurry of 87 mg (10% by weight) of 10%palladium on carbon in (5 mL) ethyl alcohol and hydrogenated for 16 hunder a slight positive pressure of hydrogen. The mixture was filteredand concentrated in vacuo to yield 553.2 mg of the title product as acolorless glass which was used directly for ensuing reaction. TLC:Rf=0.46, 10% methanol/CH₂Cl₂.

E. Compound 51. To a solution of 72.7 mg (0.207 mmol) of the resultantcompound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 22.6 mg (0.27 mmol), and2-(pyrid-2-yl)-thiophene-5-sulfonyl chloride 64.6 mg, (0.249 mmol).After 14 h, the resulting mixture was diluted with ethyl acetate, washedwith saturated brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by thin layerpreparative chromatography using 15 to 30% ethyl acetate/CH₂Cl₂ eluentto provide 53 mg of the title product as a white solid. TLC; RF=0.25,25% ethyl acetate/CH₂Cl₂, HPLC: Rt=15.3 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 52

A. N-hydroxysuccinimidyl-(RS)-3-hydroxyl-tetrahydrofuryl carbonate. Thetitle compound was prepared as described in Example 48A starting with1.0 g of (RS)-3-hydroxy-tetrahydrofuran and yielding 2.33 g of a whitesolid.

B. Compound 52. To a solution of 105 mg of the resultant compound ofExample 35A in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, 112 mg of the resultant compound of Example 52Aand 126 mg N,N-diisopropylethylamine. The mixture was stirred for 4 h,diluted with CH₂C₂, washed and saturated NaHCO₃ and saturated NaCl, thendried over MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 5% CH₃OH inCH₂Cl₂ as eluent to yield 101.4 mg of product. TLC: Rf=0.52, 5% CH₃OH inCH₂Cl₂. HPLC: Rt=15.05 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 53

Compound 53. To a solution of 72.3 mg (0.19 mmol) of the resultantcompound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 19.2 mg (0.228 mmol), and4-acetamido-3-chlorobenzene sulfonyl chloride 61.1 mg, (0.228 mmol).After 14 h, the resulting mixture was diluted with EtOAc, washed withsaturated brine, dried over magnesium sulfate, filtered and concentratedin vacuo. The residue was purified by low pressure silica gel columnchromatography using 20% to 45% EtOAc/CH₂Cl₂ eluent to provide 49.1 mgof the title product. TLC: RF=0.29, 50% EtOAc/CH₂Cl₂, HPLC: Rt=13.9 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 54

Compound 54. A solution of 260 mg of the resulting Compound of 39A and45 mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10 mL of CH₂Cl₂was reacted in the same manner as described for Example 14. After workupand purification by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 1.4 mg of thetitle compound was obtained. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC:Rt=15.63 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 55

Compound 55. 35.0 mg of the resulting compound of Example 54 was treatedwith 1 mL of 90% is aqueous TFA and allowed to stand for 12 h. Themixture was concentrated in vacuo and the residue taken up in 10 mL ofdry CH₂Cl₂, treated with 34 μL of DIEA (0.23 mmoles) and 20 mg of1-benzyl-3-tert-butyl-1H-pyrazole-5-carbonyl chloride. The mixture wasstirred for 1.5 h, then diluted with in CH₂Cl₂, and washed with 1 N HCl.After drying over MgSO₄ and concentrating in vacuo, a portion of themixture was purified by preparative reversed-phase C₁₈ HPLC using alinear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution toobtain 1.1 mg of the title compound. TLC: Rf=0.8, 5% CH₃OH in CH₂Cl₂.HPLC: Rt=18.25 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 56

A. S(−)-1-phenylethyl-N-hydroxysuccinimidyl carbonate. The titlecompound was prepared from 9.5 μL of S(−)-1-phenylethanol and 30 m ofN,N-disuccinimidyl carbonate as described in Example 44A. The resultingmaterial was used without subsequent purification; (¹H)-NMR (CDCl₃)consistent with structure.

B. Compound 56. 45.0 mg of the resulting compound of Example 58 wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. Themixture was concentrated in vacuo and the residue taken up in 15 mL ofdry CH₂Cl₂, treated with the above mixed anhydride and 65 μL oftriethylamine. The mixture was stirred for 14 h then diluted with ethylacetate and washed with saturated sodium bicarbonate solution andsaturated brine, dried over magnesium sulfate, filtered and concentratedin vacuo. A portion of the mixture was purified by preparativereversed-phase C₁₈ HPLC using a linear gradient of mixture was purifiedby preparative reversed-phase C₁₈ HPLC using a linear gradient of 35% of100% CH₃CN/H₂O with 0.1 TFA for elution to obtain 1.1 mg of the titlecompound. TLC: Rf=0.5, 3% CH₃OH in CH₂Cl₂. HPLC: Rt=17.44 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 57

Compound 57. 30 mg of the resultant compound of Example 58 was treatedwith 1 mL of 90% aqueous TFA and allowed to stand for 12 h. The mixturewas concentrated in vacuo and the residue taken up in 25 mL of dryCH₂Cl₂, washed and saturated sodium bicarbonate solution, dried overmagnesium sulfate, filtered and concentrated in vacuo. A solution of 14mg of the resultant free amine in 10 mL of CH₂Cl₂ was treated with 6 μLof phenoxyacetyl chloride and 12 μL of triethylamine. The mixture wasstirred under an inert atmosphere for 1 h, then diluted in CH₂Cl₂ andwashed with 1 N HCl. After drying over MgSO₄ and concentrating in vacuo.A portion of the mixture was purified by preparative reversed-phase C₁₈HPLC using a linear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA aseluant to obtain 16.5 mg of the title compound. TLC: Rf=0.25, 3% MeOH inCH₂Cl₂. HPLC: Rt=16.6 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 58

Compound 58. A solution of 500 mg of the resulting compound of Example39A and 370 mg of benzofurazan-4-sulfonyl chloride in 10 mL of CH₂Cl₂was reacted in the same manner as described for Example 14. Afterworkup, the title compound was obtained by crystallization from hotethanol. Further purification of this material by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluent gave 2.0 mg of the title compound. TLC: Rf=0.35,3% CH₃OH in CH₂Cl₂. HPLC: Rt=17.00 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 59

A. R(+)-1-phenylethyl-N-hydroxysuccinimidyl carbonate. The titlecompound was prepared from R(+)-1-phenylethanol as described in Example56A to yield a white solid. The resulting material was used directly forsubsequent reaction; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 59. A 36 mg portion of the resultant compound of Example 58and 0.21 μmol of the resulting compound of 59A were reacted in themanner described in example 56B. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 1.0 mg of the title compound wasobtained as a white solid. TLC: Rf=0.45, 3% MeOH in CH₂Cl₂. HPLC:Rt=17.34 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 60

Compound 60. To a solution of 70 mg of the resultant compound of Example51D in 10 mL of CH₂Cl₂ was added 3 mL of saturated aqueous sodiumbicarbonate solution, 50 mg of sodium bicarbonate, and 53 mg ofbenzofurazan-4-sulfonyl chloride. The mixture was stirred vigorously for4 h, then the resulting mixture was diluted with CH₂Cl₂, washed withsaturated brine, dried over magnesium sulfate and filtered. Afterconcentration of the mixture in vacuo, the residue was purified by thicklayer silica gel chromatography using 5% MeOH/CH₂Cl₂ as eluant to obtain80 mg of the title compound as a white solid. TLC: Rf=0.80, 5% MeOH inCH₂Cl₂. HPLC: Rt=14.96 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 61

Compound 61. To a solution of 35.5 mg (0.076 mmol) of the resultantcompound of Example 16 in 1 mL of dichloromethane was sequentially added27.6 μL (0.159 mmol) of diisopropylethyl amine and 12 μL (0.083 mmol) ofbenzyl chloroformate. After 1 hour, the mixture was concentrated invacuo. The residue was purified by preparative thin layer chromatographywith 50% ethyl acetate/dichloromethane as an eluent to yield 38.7 mg ofthe title compound as a white solid; TLC: Rf=0.63, 50% ethylacetate/dichlormethane; HPLC: Rt=15.45 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 62

A. Benzofurazan-4-sulfonic acid. To a solution of 252.0 mg (1.05 mmol)of o-nitroaniline-m-sulfonic acid sodium salt in 1 mL of water was added0.52 mL of 2.0 N HCl. After ½ h, 0.68 mL (1.05 mmol) oftetrabutylammonium hydroxide (40% in water) was added. After 2 hours,the mixture was concentrated in vacuo. A solution of the residue in 7 mLof acetic acid was treated with 488.5 mg (1.10 mmol) of leadtetraacetate. After 24 hours, the precipitate was filtered and washedwith small amount of acetic acid. The solid was further dried in vacuoto yield 267.9 mg of product. TLC: Rf=0.09, 10% CH₃OH/CH₂Cl₂.

B. Benzofurazan-4-sulfonyl chloride. To a solution of 137.0 mg (0.522mmol) of triphenylphosphine in 0.5 mL of dichloromethane was slowlyadded 47 μL (0.594 mmol) of sulfuric chloride at 0° C. The ice-waterbath was removed and the crude resultant compound of Example 62A in 0.5mL of dichloromethane was added slowly. After 3 hours, the mixture wastreated with 30 mL of 50% ether/hexane. The supernatant was decantedinto a dry flask and concentrated in vacuo. The residue was purified byfiltering through a plug of silica gel with 25% ethyl acetate as aneluent to yield 23 mg of product. TLC: Rf=0.6, 10% CH₃OH/CH₂Cl₂;(¹H)-NMR (CDCl₃) consistent with structure.

C. Compound 62. To a solution of 55.7 mg (0.166 mmol) of the resultantcompound of Example 39A in 1 mL of dichloromethane was sequentiallyadded 0.5 mL of saturated NaHCO₃, a small amount of solid NaHCO₃ and theresultant compound of Example 62B. After 3 hours, the mixture wasdiluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 5.3 mg of the title compound as a white solid; TLC: Rf=0.40, 50%ethyl acetate/dichloromethane; HPLC Rt=16.5 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 63

A. A solution of 3.0 mg (0.0058 mmol) of the resultant title compound ofExample 62 in 2 mL ethyl acetate was treated with HCl gas (moderatestream) for 3 minutes. The mixture was concentrated in vacuo to yieldthe crude amine hydrochloride salt. TLC: Rf=0.20, 10% CH₃OH/CH₂Cl₂.

B. Compound 63. To a solution of the crude resultant compound of Example63A in 1 mL of dichloromethane was sequentially added 2.1 uL (0.0121mmol) of diisopropyl ethyl amine and 0.9 uL (0.0064 mmol) of benzylchloroformate. After 1 hour, the mixture was concentrated in vacuo. Theresidue was purified by preparative thin layer chromatography with 90%dichloromethane/methanol as an eluent to yield 2.6 mg of the titlecompound as a white solid; TLC: Rf=0.34, 50% ethylacetate/dichloromethane; HPLC, Rt=17.1 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 64

A. 5-(Dimethylamino)thioxomethoxy)-benzofurazan. To a solution of 500 mg(3.67 mmol) of 5-hydroxybezofurazan in 10 mL of DMF was added 140 mg(4.59 mmol) of NaH in small portions. The resulting mixture was stirredat room temperature until no more gas evolved. The flask was thenimmersed in a cold water bath and 540 mg (4.41 mmol) ofdimethylthiocarbamoyl chloride (from Aldrich) was added. After 5minutes, the water bath was removed the mixture was heated to 80° C. for1 hour. After being cooled to room temperature, the mixture was pouredinto 20 mL of 0.5 N NaOH three times and water three times. The solidwas dried in vacuum to yield 580 mg of product that was used in the nextreaction without further purification; TLC: Rf=0.20, 20% ethylacetate/hexane; (¹H)-NMR (CDCl₃) consistent with structure.

B. 5-((Dimethylamino)carbonyl)thio)-benzofurazan. The crude product, 510mg (2.28 mmol), from Example 64A was heated to 190° C. in a sealed tube.After 5 hours, it was cooled to room temperature and ethyl acetate wasadded. The solution was filtered through a plug of a silica andconcentrated in vacuo to yield 360 mg of product which was again used inthe next reaction without further purification. TLC: Rf=0.20, 20% ethylacetate/hexane.

C. 5-Mercaptobenzofurazan. To a solution of 357.4 mg (1.60 mmol) of theresultant compound of Example 64B in 2 mL of methanol was added 7 mL of6 N NaOH. The mixture was heated to 90° C. for 2 hours. The mixture waspoured into 100 mL ice and acidified with concentrated HCl. The slurrywas filtered and rinsed three times with water. The residue was dried invacuo to yield 145.6 mg of product; TLC: Rf=0.70, 20 ethylacetate/hexane; (¹H)-NMR (CDCl₃) consistent with structure.

D. Benzofurazan-5-sulfonyl chloride. Chlorine gas was bubbled through asolution of 39.9 mg (0.26 mmol) of the resultant compound of Example 64Cin a mixture of 1 mL of ethyl acetate and 0.5 mL of water for 3 minutes.The mixture was then washed repeatedly with brine until no moreprecipitate formed. The organic layer was dried over MgSO₄, filtered andconcentrated to yield 30 mg of the product (52%). TLC: Rf=0.22, 20%ethyl acetate/hexane.

E. Compound 64. A solution of the resultant compounds of Examples 52Dand 39A (total yields) in a mixture of 1 mL of dichloromethane, 0.3 mLof saturated NaHCO₃ and a small amount of solid NaHCO₃ was stirred atroom temperature for 2 hours. The solution was diluted with 30 mL ofdichloromethane and the two layers were separated. The aqueous layer wasextracted once with dichloromethane chloride. The combined organic layerwas washed with brine, dried over MgSO₄ and concentrated. The residuewas purified by preparative thin layer chromatography with 90%dichloromethane/ether as an eluent to yield 30 mg of the title productas a white solid; TLC: Rf=0.46, 10% Et₂O/CH₂Cl₂, HPLC Rt=17.6 min;(¹H)-NMR (CDCl₃): δ8.45 (s), 1H, 7.96 (d), 1H, 7.65 (d), 1H, 7.25 (m),5H, 4.65 (d), 1H, 3.85 (m), 1H, 3.78 (m), 1H; 3.30 (d), 2H; 3.10 (m),2H, 2.90 (m), 2H, 1.90 (m), 1H; 1.40 (s), 9H; 0.90 (d), 6H.

EXAMPLE 65

Compound 65. A solution of 13.1 mg (0.025 mmol) of the resultantcompound of Example 64E in 1.5 mL of ethyl acetate was treated withgaseous HCl (moderate stream) at 0° C. for 3 minutes. The solvent wasremoved to yield a solid residue which was used in the next reactionwithout further purification; TLC: Rf=0.52, 10% CH₃OH/CH₂Cl₂. A solutionof this hydrochloride salt (entire yield) in 1 mL of dichloromethane wastreated sequentially with 9.2 μL (0.053 mmol) of diisopropyl ethyl amineand 4.0 μL (0.028 mmol) of benzyl chloroformate. After 3 hours, themixture was concentrated and purified by preparative thin layerchromatography with 90% dichloromethane/ether as an eluent to yield 11.7mg of the title compound as a white solid; TLC: Rf=0.65, 10%Et₂O/CH₂Cl₂; HPLC Rt=17.6 min; (¹H)-NMR (CDCl₃: δ 8.45 (s), 1H, 7.96(d), 1H, 7.65 (d), 1H, 7.25 (m), 10H, 5.00, (m), 2H, 4.85 (d), 1H, 3.86(m), 2H, 3.60 (bs), 1H, 3.25 (m), 12H, 3.05 (d), 2H, 2.96 (m), 1H, 2.98(m), 1H, 1.88 (m), 1H, 0.90 (dd), 6H.

EXAMPLE 66

Compound 66. A solution of 100 mg (0.46 mmol) of the resultant compoundof Example 64D and 101 mg (0.286 mmol) of the resultant compound ofExample 48A in a mixture of 2 mL of dichloromethane, 0.5 mL of saturatedNaHCO₃ and small amount of solid NaHCO₃ was stirred at room temperaturefor 2 hours. The solution was diluted with 50 mL of dichloromethane andthe two layers were separated. The aqueous layer was extracted once withdichloromethane. The combined organic layer was washed with brine, driedover MgSO₄ and concentrated. The residue was purified by preparativethin layer chromatography with 20% ethyl acetate/hexane as an eluent toyield 82 mg of the title product as a slightly impure pale yellow solid.The material was further purified by preparative HPLC with a lineargradient solvent system of 35% to 80% of acetonitrile/water (0.1% TFA)over 80 min. Upon removal the solvents 50 mg of white solid wasobtained. TLC: Rf=0.46, 10% Et₂O/CH₂Cl₂; HPLC, Rt=17.6 min; (¹H)-NMR(CDCl₃): δ 8.45 (s), 1H, 7.96 (d), 1H, 7.65 (d), 1H, 7.25 (m), 5H, 5.15(m), 1H, 4.85 (d), 1H, 3.82 (m) 4H, 3.68 (d), 1H, 3.20 (m), 2H, 3.05(d), 2H, 2.96 (m), 1H; 2.88 (m), 1H, 2.14 (m), 1H, 1.92 (m), 2H, 1.50(bs), 1H, 0.90 (dd), 6H.

EXAMPLE 67

Compound 67. Following the procedure described in Example 40B, asolution of the resultant compound of Example 40A in CH₂Cl₂ is treatedwith bis-((carboxamido)-amino)-acetic acid, diisopropylethylamine, HOBt,and EDC in a 1:1:1:1:1 molar ratio. the mixture is stirred for 16 h atambient temperature while protected from moisture, then diluted withadditional CH₂Cl₂ and washed sequentially with H₂O, saturated NaHCO₃solution and brine, then dried over MgSO₄ and concentrated in vacuo. Theresidue is purified by silica gel chromatography using an appropriateeluant to yield the title product.

EXAMPLE 68

Compound 68. This compound was prepared by the route described inExample 26, except that the reacting amine used was the resultingcompound of Example 39A (146 mg, 0.43 mmol) and the acylating agent was4-fluorophenyl sulphonyl chloride (27 mg, 0.14 mmol). Afterchromatographic purification on a silica gel column using 8%CH₃OH/CH₂Cl₂ as eluent, 92.8 mg of the title compound was obtained.HPLC: Rt=15.9 minutes. TLC: Rf=0.54, 8% MeOH/CH₂Cl₂; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 69

A. The resulting compound of Example 68 (72.1 mg, 0.167 mmol) wasdissolved in 90% aqueous TFA (3.3 mL), and the reaction mixture stirredfor 3 hours at room temperature, then was concentrated to dryness. TLC:Rf=0.29, 8% MeOH/CH₂Cl₂.

B. Compound 69. To a solution of the resulting compound of Example 69A(41.7 mg, 0.09 mmol) in CH₂Cl₂ (2 mL) was added diisopropylethylamine(47 μl, 0.27 mmol) and the resulting compound of Example 48A (33 mg,0.15 mmol), and the reaction proceeded for 14 hours at room temperature.The reaction mixture was then concentrated, and the residue waschromatographed on a silica gel column using 8% THF/CH₂Cl₂ was eluent,yielding the desired compound which was further subjected topurification by preparative HPLC, yielding 7.8 mg of a white solid.HPLC: Rt=13.5 minutes. TLC: Rf=0.36, 8% THF/CH₂Cl₂; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 70

Compound 70. A solution of 30 mg of the resulting compound of Example 54and 17.6 mg of 3-acetamido-4-fluorobenzenesulfonyl chloride in 10 mL ofCH₂Cl₂ was reacted in the same manner as described for Example 14. Afterworkup and purification by preparative reversed-phase Cl₈ HPLC using alinear gradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluent, 2.0 mgof the title compound was obtained. TLC: Rf=0.5, 10% CH₃OH in CH₂Cl₂.HPLC: Rt=13.74 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 71

Compound 71. A 30 mg portion of the resultant compound of Example 58 wasdeprotected with trifluoroacetic acid and the resulting compound reactedwith 9 μL of dimethysulfamoyl chloride in 10 mL of CH₂Cl₂ was reacted inthe manner described in Example 14. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 6.5 mg of the title compound wasobtained. TLC: Rf=0.2, 3% MeOH in CH₂Cl₂. HPLC: Rt=15.96 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 72

Compound 72. A solution of the resulting compound from thetrifluoroacetic acid deprotection of Example 69A (31 mg, 0.07 mmol) inCH₂Cl₂ (2 mL) was added diisopropylethylamine (47 μl, 0.27 mmol) anddimethylsulfamoyl chloride (22 μl, 0.20 mmol), and the reactionproceeded for 16 hours at room temperature. The reaction mixture wasthen concentrated, and the residue was chromatographed on a thick layersilica gel plate (1.0 mm) using 5% THF/CH₂Cl₂ as eluent, yielding thedesired compound which was further subjected to purification bypreparative HPLC to yield 7.8 mg of a white solid. HPLC: Rt=14.8minutes. TLC: Rf=0.44, 5% THF/CH₂Cl₂.

EXAMPLE 73

Compound 73. A 43 mg portion of the resultant compound of Example 54 wastreated with 1 mL of 90% aqueous TFA and allowed to stand for 12 h. Themixture was concentrated in vacuo and the residue taken up in 5 mL ofCH₂Cl₂. To this solution, 3 mL saturated aqueous sodium bicarbonate and25 mg of 2,5-dimethoxybenzenesulfonyl chloride was added, and themixture was stirred for 12 h, warming slowly to ambient temperature.After concentration of the mixture in vacuo, the residue was purified bythick layer silica gel chromatography using 3% MeOH/CH₂Cl₂ as eluantfollowed by preparative reversed-phase C₁₈ HPLC using a linear gradientof 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 5.5 mg of thetitle compound. TLC: Rf=0.20, 3% MeOH/CH₂Cl₂. HPLC: Rt=15.15 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 74

A. Compound XXI (A=tert-butoxycarbonyl, D′=cyclopropylmethyl, A′=H). Toa solution of compound XX (A=tert-butoxycarbonyl)(0.8 g, 2.67 mmol) inethanol (30 mL) was added a solution of KOH (0.18 g, 3.2 mmol) inethanol (20 mL) and the mixture stirred for 45 min at room temperature.In a separate flask, a solution of cyclopropylmethyl-amine hydrochloride(1.44 g, 13.3 mmol) in ethanol (20 mL) was added KOH (0.75 g, 13.3mmol). The mixture was stirred 30 min at room temperature. The solutionswere combined and heated at 85° C. for 3 h. The solution wasconcentrated under reduced pressure and the residue slurried in diethylether and filtered. The ethereal layer was concentrated to give 0.32 gof a white solid; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 74. To a solution of the resulting compound of Example 74A(0.1 g, 0.30 mmol) in CH₂Cl₂ (20 mL) was added a saturated solution ofsodium bicarbonate, followed by addition of solid sodium bicarbonate (30mg, 0.36 mmol), then 4-fluorobenzenesulfonyl chloride (0.07 g, 0.36mmol). The mixture was allowed to stir at room temperature for 4 h. Theorganics were extracted into 250 mL CH₂Cl₂, dried over anhydrous MgSO₄,concentrated under reduced pressure then purified via medium pressureliquid chromatography using a gradient system of CH₂Cl₂ followed by0.5:99.5 methanol/CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂. The titlecompound was obtained as 35 mg of a colorless foam. HPLC: Rt=16.8 min.TLC: Rf=0.32, 3:97 methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 75

A. Compound XXI (A=tert-butoxycarbonyl, D′=isopropyl, A′=H). To asolution of Compound XX (A=tert-butoxycarbonyl)(1.67 mmol) in ethanol(10 mL) was treated with isopropylamine (10 mL). The solution was heatedto 85° C. for 72 h. The solution was filtered then concentrated underreduced pressure to give 0.56 g of the title compound which was usedwithout-subsequent purification. (¹H)-NMR (CDCl₃) consistent withstructure.

B. Compound 75. To a solution of the resultant compound of Example 75A(0.2 g, 0.65 mmol) in CH₂Cl₂ (10 mL) was added a saturated solution ofsodium bicarbonate (3 mL), followed by addition of solid sodiumbicarbonate (0.11 g, 1.31 mmol), then p-fluorobenzenesulfonyl chloride(0.25 g, 1.28 mmol). The mixture was stirred overnight at ambienttemperature. The organics were extracted into 100 mL CH₂Cl₂, dried overanhydrous MgSO₄, concentrated under reduced pressure then purified viamedium pressure silica gel chromatography using a gradient system ofCH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂. The title compound was obtainedas a colorless foam 200 mg. TLC: Rf=0.22, 3:97 methanol/CH₂Cl₂, HPLC:Rt=16.48 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 76

A. Compound XXI (A=tert-butoxycarbonyl, D′=morpholinyl, A′=H). To asolution of compound XX (A=Boc) in ethanol is added 3 molar equivalentsof N-amino morpholine. The mixture is heated under reflux for 12 h,cooled, and the mixture concentrated in vacuo. The residue is purifiedby preparative reversed-phase chromatography using a linear gradient of5% to 100% acetonitrile/H₂O as eluant to yield the title compound.

B. Compound 76. Following the procedure described in Example 81, asolution of the resultant compound of Example 76A in CH₂Cl₂ is reactedwith 4-fluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄, filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 77

A. Compound XXI (A=tert-butoxycarbonyl, D′=4-(N,N-dimethylamino)-benzyl,A′=H). To a solution of compound XX (A=Boc) in ethanol is added 3 molarequivalents of 4-aminomethyl-(N,N-dimethyl)-aniline. The mixture isheated under reflux for 12 h, cooled, and the mixture concentrated invacuo. The residue is purified by silica gel chromatography using anappropriate solvent system as eluant to yield the title product.

B. Compound 77. Following the procedure described in Example 81, asolution of the resultant compound of Example 77A in CH₂Cl₂ is reactedwith 4-fluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄, filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 78

A. Compound XXI (A=tert-butoxycarbonyl, D′=cyclopentyl, A′=H). To asolution of compound XX (A=Boc) in ethanol is added 10 molar equivalentsof cyclopentylamine. The mixture is heated under reflux for 12 h,cooled, and the mixture concentrated in vacuo. The residue is usedwithout subsequent purification.

B. Compound 78. Following the procedure described in Example 81, asolution of the resultant compound of Example 78A in CH₂Cl₂ is reactedwith 4-fluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄, filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 79

A. Compound XXI (A=tert-butoxycarbonyl, D′=2-(4-pyridyl)ethyl, A′=H). Toa solution of compound XX (A=Boc) in ethanol is added 3 molarequivalents of 4-aminoethylpyridine. The mixture is heated under refluxfor 12 h, cooled, and the mixture concentrated in vacuo. The residue ispurified by preparative reversed-phase chromatography using a lineargradient of 5% to 100% acetonitrile/H₂O as eluant to yield the titleproduct.

B. Compound 79. Following the procedure described in Example 81, asolution of the resultant compound of Example 79A in CH₂Cl₂ is reactedwith 4-fluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄, filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography usingan appropriate solvent system to yield the title product.

EXAMPLE 80

A. 4-Cyanotetrahydro-4H-pyran. Following essentially the procedure ofYoneda, R. “Cyanophosphate: An Efficient intermediate for Conversion ofCarbonyl compounds to Nitriles,” Tetrahedron Lett., 30, 3681 (1989), asolution of tetrahydro-4H-pyran-one (9.9 g, 97.8 mmol) in dry THF (50mL) is reacted with lithium cyanide (9.7 g, 294 mmol) anddiethylcyanophosphonate (24 g, 146 mmol). The mixture is stirred for 24h at ambient temperature. The reaction is quenched by the addition of100 mL H₂O. The product is extracted into 1.5 L of diethyl ether, driedover anhydrous MgSO₄ then concentrated under reduced pressure. Theresidue is dissolved in dry THF (30 mL) and tert-butyl alcohol (7.25 g,97.8 mmol). This solution is added slowly to 75 mL of a 1 M solution ofSmI₂. The mixture is stirred for 15 h at ambient temperature. Thereaction is quenched by addition of 100 mL of saturated aqueous NH₄Cl.The resulting mixture is extracted with diethyl ether and the organiclayers dried over anhydrous MgSO₄ and concentrated under reducedpressure. Purification by silica gel chromatography gives the titlecompound.

B. 4-(aminomethyl)tetrahydro-4H-pyran To a solution of the compound ofthe Example 80A (10 g, 89.9 mmol) in absolute ethanol (200 mL) is addedRaney Nickel (2.0 g, 50% slurry in water). The mixture is stirred for 24hours at ambient temperature under 40 psig of hydrogen. The solution isfiltered through celite and the solution concentrated under reducedpressure. The residue is taken up in ether (2L) washed with brine, driedin anh. MgSO₄, then concentrated under reduced pressure to give thetitle compound.

C.(1S,2R)-N-(1-Benzyl-3-(N-(4-(aminomethyl)tetrahydro-4H-pyran))-2-hydroxypropyl)-tertbutoxycarbonylamine.

To a solution of the compound of Example 80B (5 g, 48.5 mmol) inabsolute ethanol (20 mL) is added the compound XX (A=Boc)(2.55 g, 9.7mmol). The mixture is stirred for 24 hours at ambient temperature. Thesolution is concentrated under reduced pressure and the crude product ispuffed via column chromatography to give the title compound.

D. Compound XXII (A=Boc, D′=(4-tetrahydro-4H-pyranyl)methyl, A′=H). To asolution of compound XX (A=Boc) in ethanol is added 3 molar equivalentsof the resulting compound of Example 80C. The mixture is heated underreflux for 12 h, cooled, and the mixture concentrated in vacuo. Theresidue is purified by preparative reversed-phase chromatography using alinear gradient of 5% to 100% acetonitrile/H₂O as eluant to yield thetitle compound.

To a solution of compound XX (A=Boc) in ethanol is added 3 molarequivalents of N-amino morpholine. The mixture is heated under refluxfor 12 h, cooled, and the mixture concentrated in vacuo. The residue ispurified by preparative reversed-phase chromatography using a lineargradient of 5% to 100% acetonitrile/H₂O as eluant to yield the titlecompound.

E. Compound 80. Following the procedure described in Example 81, asolution of the resultant compound of Example 80D in CH₂Cl₂ is reactedwith 4-fluorobenzenesulfonyl chloride in the presence of water andNaHCO₃. Following dilution with additional CH₂Cl₂ and aqueous workup,the resultant product is dried over MgSO₄, filtered, and concentrated invacuo. The residue is then purified by silica gel chromatography toyield the title product.

EXAMPLE 81

A. Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,E=3,4-dichlorophenyl). A solution of 316 mg of the resultant compound ofExample 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treatedsequentially, at ambient temperature under an atmosphere of nitrogen,with 276 mg of 3,4-dichorobenzenesulfonyl chloride and 95 mg of sodiumbicarbonate. The mixture was stirred for 14 h, diluted with CH₂Cl₂,washed with saturated NaCl then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 5% diethyl ether/CH₂Cl₂ as eluent to yield 490mg of product. TLC: Rf=0.26, 5% diethyl ether in CH₂Cl₂. HPLC: Rt=18.92min. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (P═H, D′=isobutyl, E=3,4-dichlorophenyl, hydrochloridesalt). A solution of 467 mg of the resultant compound of Example 81A inethyl acetate was treated at −20° C. with HCl gas. The HCl was bubbledthrough the mixture for 20 min over which time the temperature wasallowed to warm to 20° C. Nitrogen was then bubbled through the mixturefor 15 min and solvent removed in vacuo to yield 412 mg of product as awhite solid which was used without subsequent purification.

C. Compound 81. A solution of 91 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 25 mg of allyl chloroformate and 52 mgN,N-diisopropylethylamine. The mixture was stirred for 4 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 89 mg of the title productas a white solid. TLC: Rf=0.53, 5% diethyl ether in CH₂Cl₂. HPLC:Rt=17.95 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 82

A. (3-Pyridyl)-methyl-4-nitrophenyl-carbonate. To a solution of 3.65 gof bis-(nitrophenyl) carbonate in 25 mL of CH₂Cl₂ at 0° C. was addedsequentially 0.97 mL of 3-pyridyl carbinol and 1.3 mL of 4-methylmorphine. After stirring at room temperature for 24 hours, the resultantmixture was diluted with 100 mL of CH₂Cl₂, washed with saturated sodiumbicarbonate, water and brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by filtration through aplug of silica gel, using 0-40% EtOAc/CH₂Cl₂ as eluent to provide 1.68 gof the title product. TLC: Rf=0.19, 50% EtOAc/hexane.

B. Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,E=3,4-benzofurazan). To a solution of 498.6 mg of the resultant compoundof Example 39A in 10 mL of CH₂Cl₂ was added sequentially, 2 mL ofsaturated sodium bicarbonate, a small amount of solid sodium bicarbonateand 518.4 mg of the resultant compound of Example 64D. After stirring atroom temperature for 3 hours, the resultant mixture was diluted with 60mL of CH₂Cl₂, washed with saturated sodium bicarbonate and brine, driedover magnesium sulfate, filtered and concentrated in vacuo. The residuewas purified by silica gel chromatography using 5% diethyl ether/hexaneas eluent to yield 300 mg of white solid. TLC: Rf=0.80, 50%EtOAc/hexane.

C. Compound XXII (P═H, D′=isobutyl; E=3,4-benzofurazan, hydrochloridesalt.). A solution of 60.3 mg of the resultant compound of Example 82Bin 3 mL of EtOAc at −20° C. was treated with anhydrous HCl gas for 5min. The ice bath was removed and after an additional 10 min. Thereaction mixture was sparged with nitrogen then concentrated in vacuumand the resulting white solid used without subsequent purification forsubsequent reaction.

D. Compound 82. To a solution of the resultant compound of Example 82C(entire yield) in 2 mL of CH₂Cl₂ was added sequentially, 45 μL ofdiisopropylethylamine and 35.1 mg of the resultant compound of Example82A. The mixture was stirred for 24 hours and then concentrated invacuo. The residue was purified by preparative thin layer chromatographyusing 60% ether/CH₂Cl₂ as eluent followed by preparative reversed-phaseC₁₈ HPLC using a linear gradient of 40% to 100% CH₃CN/H₂O with 0.1% TFAas eluant. The resultant TFA salt of the title compound was washed withsaturated sodium bicarbonate to yield 6.5 mg of the title compound. TLC:Rf=0.15, 20% EtOAc/CH₂Cl₂. HPLC: Rt=13.52 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 83

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=4-acetamido-3-chlorophenyl). A solution of 339 mg of the resultantcompound of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ wastreated sequentially, at ambient temperature under an atmosphere ofnitrogen, with 324 mg of 4-acetamido-3-chlorobenzenesulfonyl chlorideand 102 mg of sodium bicarbonate. The mixture was stirred for 14 h,diluted with CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 20% diethyl ether in CH₂Cl₂ aseluent to yield 498 mg of product. TLC: Rf=0.27 (20% diethyl ether inCH₂Cl₂). HPLC: Rt=16.20 min. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=isobutyl, E=4-acetamido-3-chlorophenyl,hydrochloride salt). A solution of 474 mg of the resultant compound ofExample 83A in ethyl acetate was treated at −20° C. with HCl gas. TheHCl was bubbled through the mixture for 20 min over which time thetemperature was allowed to warm to 20° C. Nitrogen was then bubbledthrough the mixture for 15 min and the solvent was removed in vacuo toyield 421 mg of product as a white solid which was used withoutsubsequent purification.

C. Compound 83. A solution of 92 mg of the resultant compound of Example83B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 24 mg of allyl chloroformate and 52 mgN,N-diisopropylethylamine. The mixture was stirred for 4 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl, then dried over MgSO₄,filtered, and concentrated in vacuo to yield 106 mg of the title productas a white solid. TLC: Rf=0.38 (20% diethyl ether in CH₂Cl₂). HPLC:Rt=15.28 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 84

Compound XXII (P=tert-butoxycarbonyl, D′=isobutyl,E=3,4-dichlorophenyl). To a solution of the resultant compound ofExample 51D (220 mg, 0.61 mmol) in CH₂Cl₂ (10 mL) was added the3,4-dichlorobenzenesulfonyl chloride (300 mg, 1.22 mmol) followed by theaddition of a saturated solution of sodium bicarbonate (3 mL) followedby addition of 0.1 g of solid sodium bicarbonate. The mixture wasstirred at ambient temperature overnight. The solution was diluted with100 mL CH₂Cl₂, the organics separated, dried over anhydrous, MgSO₄, andthe organics concentrated under reduced pressure to obtain 0.17 g ofcrude product. The crude product was purified via medium pressure liquidchromatography using CH₂Cl₂ followed by 0.5:99.5 methanol/CH₂Cl₂followed by a 1:99 methanol/CH₂Cl₂ solution as the solvent system togive 103 mg of the title compound as a white solid. TLC: Rf=0.56 (3:97methanol/CH₂Cl₂), HPLC: Rt=19.78 min, (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 85

A. (3-Tetrahydrofuryl)-methyl-4-nitrophenyl-carbonate. To a solution of1.21 g of p-nitrophenyl chloroformate in 20 mL of CH₂Cl₂ 0° C. was addedsequentially, 0.51 g of tetrahydro-3-furanmethanol and 0.66 mL of4-methyl morpholine. After stirring at room temperature for 2 hours. Themixture was stirred for 2 hours and concentrated in vacuo. The residuewas purified by filtering through a plug of silica gel, using 0-50%EtOAc/CH₂Cl₂ as eluent to provide 1.17 g of the title product as a paleyellow solid. TLC: Rf=0.20, 50% EtOAc/hexane.

B. Compound 85. To a solution of 70 mg of the resultant compound ofExample 81B in 1 mL of THF was added sequentially, 56 μL ofdiisopropylethylamine and a solution of 46.6 mg of the resultantcompound of Example 85A in 1 mL of THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was diluted with 60 mLof CH₂Cl₂, washed with 5% sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated in vacuo to yield 120 mg ofcrude product. The residue was purified by preparative thin layerchromatography using 20% EtOAc/CH₂Cl₂ as eluent to yield 82 mg of thetitle compound. TLC: Rf=0.4, 20% EtOAc/CH₂Cl₂. HPLC: Rt=17.08 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 86

Compound 86. A solution of 42 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 41 mg of the product of Example 52A and 46mg N,N-diisopropylethylamine. The mixture was stirred 14 h, diluted withCH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then dried overMgSO₄, filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using ethyl acetate aseluent to yield 43 mg of product. TLC: Rf=0.44 (20% ethyl acetate).HPLC: Rt=13.14 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 87

A. Compound XXII (P═H, D′=isobutyl, E=4-acetamido, 3-fluoro). A solutionof 25 mg of the resultant compound of Example 54 in EtOAc (10 mL) at 0°C. was treated with anhydrous hydrogen chloride gas for 10 min., andallowed to stand for 12 h while warming to ambient temperature. Theresultant mixture was then concentrated in vacuo to yield compound as awhite solid which was used without subsequent purification for ensuingreaction.

B. Compound 87. A 0.045 mmol portion of the resultant compound ofExample 87A was taken up in 5 mL of CH₂Cl₂. To this solution, 40 μL ofdiisopropylethylamine and 6 μL of allyl chloroformate were added at 0°C. and the mixture was stirred for 12 h, while warming slowly to ambienttemperature. The resulting mixture was diluted with CH₂Cl₂, washed withsaturated brine, dried over magnesium sulfate and filtered. Afterconcentrated in vacuo, the residue was purified by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1 TFA as eluant to obtain 11.6 mg of the title compound. TLC:Rf=0.20, 5% MeOH/CH₂Cl₂. HPLC: Rt=14.6 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 88

Compound 88. A 0.033 mmol portion of the resultant compound of Example87A was taken up in 5 mL of CH₂Cl₂. To this solution, 26 μL oftriethylamine and 12 mg of the resultant compound of Example 48A wereadded and stirred for 12 h. The resulting mixture was diluted withCH₂Cl₂, washed with saturated sodium bicarbonate solution and saturatedbrine, dried over magnesium sulfate and filtered. After concentration ofthe mixture in vacuo, the residue was purified by thick layer silica gelchromatography using 5% MeOH/CH₂Cl₂ as =eluant followed by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluant to obtain 7.5 mg of the title compound. TLC:Rf=0.30, 5% MeOH/CH₂Cl₂. HPLC: Rt=13.38 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 89

Compound 89. A solution of 28 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of n-propyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 31 mg of the title productas a white solid. TLC: Rf=0.35 (5% diethyl ether in CH₂Cl₂). HPLC:Rt=18.12 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 90

Compound 90. A solution of 28 mg of the resultant compound of Example83B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 7 mg of n-propyl chloroformate and 15 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 30 mg of the title productas a white solid. TLC: Rf=0.47 (20% diethyl ether in CH₂Cl₂). HPLC:Rt=15.41 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 91

A. 3-Acetamidobenzene sulfonic acid. A solution of 1.48 g of3-aminobenzene sulfonic acid in 1:1 tetrahydrofuran/water was treated at0° C. with 1.43 g of sodium bicarbonate. After 5 min, 1.30 g of aceticanhydride was added dropwise and the reaction allowed to warm to ambienttemperature under an atmosphere of nitrogen over 14 h. The reactionmixture was passed through a column of Amberlyst 15 ion exchange resin,eluted with water, and concentrated in vacuo to yield an oil which upontreatment with benzene and azeotropic removal of water in vacuo yielded1.8 g of the title product as a white crystalline solid. (¹H)-NMR(CDCl₃) consistent with structure.

B. 3-Acetamidobenzene sulfonic acid, sodium salt. The resultant compoundof Example 91A in water was treated at 0° C. with 8.5 mL of 1 N sodiumhydroxide. The mixture was stirred for 3 h and concentrated in vacuo toyield an oil which upon treatment with benzene and azeotropic removal ofwater in vacuo yielded the title product as a tan solid which was useddirectly in the next reaction.

C. 3-Acetamidobenzenesulfonyl chloride. The resultant compound ofExample 91B in CH₂Cl₂ was treated at 0° C. with 4.5 g of phosphorouspentachloride under an atmosphere of nitrogen. The mixture was stirred14 h, extracted with CH₂Cl₂, and concentrated in vacuo to yield 1.7 g ofthe title product as a brown oil. TLC: Rf=0.21 (1:1 toluene/diethylether). (¹H)-NMR (CDCl₃) consistent with structure.

D. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=3-acetamidophenyl). A solution of 280 mg of the resultant compound ofExample 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treatedsequentially, at ambient temperature under an atmosphere of nitrogen,with 252 mg of the resultant compound of Example 91C and 105 mg ofsodium bicarbonate. The mixture was stirred for 60 h, diluted withCH₂Cl₂, washed with saturated NaCl then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 20% diethyl ether in CH₂Cl₂ as eluent to yield156 mg of the title product. TLC: Rf=0.14 (20% diethyl ether in CH₂Cl₂was). HPLC: Rt=15.39 min. (¹H)-NMR (CDCl₃) consistent with structure.

E. Compound XXII (A=H, D′=isobutyl, E=3-acetamidophenyl, hydrochloridesalt). A solution of 123 mg of the resultant compound of Example 91D inethyl acetate was treated at −20° C. with HCl gas. The HCl was bubbledthrough the mixture for 20 min, over which time the temperature wasallowed to warm to 20° C. Nitrogen was then bubbled through the mixturefor 15 min and solvent removed in vacuo to yield 118 mg of the titleproduct as a white solid which was used directly in subsequentreactions.

F. Compound 91. A solution of 49 mg of the resultant compound of Example91E in CH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, to a solution of 48 mg of the resultant compound of Example48A and 54 mg N,N-diisopropylethylamine in CH₂ Cl₂. The mixture wasstirred for 14 h, diluted with CH₂Cl₂, washed with saturated NaHCO₃ andsaturated NaCl, then dried over MgSO₄, filtered and concentrated invacuo. The residue was subjected to preparative thin layer silica gelchromatography using 5% CH₃OH in CH₂Cl₂ to yield 42 mg of product. TLC:Rf=0.32 (5% CH₃OH in CH₂Cl₂). HPLC: Rt=13.27 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 92

Compound 92. To a solution of 63.5 mg of the resultant compound ofExample 17B, diastereomer B in 1 mL of THF was added sequentially, 52 μLof diisopropylethylamine and a solution of 43.3 mg of the resultantcompound of Example 85A in 1 mL of THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was diluted with 60 mLof CH₂Cl₂, washed with 5% sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated in vacuo to yield 70.7 mgof crude product. The residue was purified by preparative reversed-phaseC₁₈ HPLC using a linear gradient of 30% to 100% CH₃CN/H₂O with 0.1% TFAas eluant to obtain 43.9 mg of the title compound. TLC: Rf=0.29, 100%EtOAc. HPLC: Rt=13.24 min; (1H) NMR (CDCl₃) consistent with structure.

EXAMPLE 93

A. N-hydroxysuccinimidyl-(R)-3-hydroxytetrahydrofuryl carbonate. Thetitle compound was prepared as described in Example 48A starting with 81mg of (R)-3-hydroxytetrahydrofuran to yield 56 mg of the title productas a white solid. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 93. To a solution of 43 mg of the resultant compound ofExample 35A in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, 27 mg of the resultant compound of Example 93Aand 39 mg N,N-diisopropylethylamine. The mixture was stirred for 14 h,diluted with CH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl,then dried over MgSO₄, filtered, and concentrated in vacuo. The residuewas purified by preparative thin layer silica gel chromatography using2% CH₃OH in CH₂Cl₂ as eluent to yield 45 mg of the title product as awhite solid. TLC: Rf=0.52 (5% CH₃OH CH₂Cl₂). HPLC: Rt=14.94 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 94

Compound 94. A solution of 47 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 28 mg of the product of Example 93A and 39mg N,N-diisopropylethylamine. The mixture was stirred for 14 h, dilutedwith CH₂Cl₂, washed with saturated NaHCO₃ and saturated NaCl, then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by preparative thin layer silica gel chromatography using 5%methanol in CH₂Cl₂ as eluent to yield 40 mg of the title product as awhite solid. TLC: Rf=0.38 (ethyl acetate). HPLC: Rt=13.09 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 95

Compound 95. To a solution of 72.0 mg (0.189 mmol) of the resultantcompound of Example 51D in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 19.1 mg (0.227 mmol), and2,3-dichlorothiophenesulfonyl chloride 57.1 mg, (0.227 mmol). After 14h, the resulting mixture was diluted with EtOAc, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by low pressure silica gel columnchromatography using 5 to 12% EtOAc/CH₂Cl₂ eluent to provide 49.1 mg ofthe title product. TLC: Rf=0.62 25% EtOAc/CH₂Cl₂, HPLC: Rt=17.3 min;(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 96

A. (4-Acetamido)-phenylmethyl-4-nitrophenyl-carbonate. To a solution of242.8 mg of p-nitrophenyl chloroformate in 5 mL of acetonitrile at 0° C.was added sequentially, 165.2 mg of 4-acetamidobenzyl alcohol and 0.13mL of 4-methyl morpholine. The mixture was stirred for 24 hours andconcentrated in vacuo. The residue was taken up in CH₂Cl₂ and washedwith 5% sodium bicarbonate and brine, dried over magnesium sulfate,filtered and concentrated in vacuo to yield 320 mg of the titlecompound. TLC: Rf=0.23, 50% EtOAc/hexane.

B. Compound 96. To solution of the resultant compound of Example 40A in1 mL of THF was added sequentially, 56 μL of diisopropylethylamine and63 mg of the resultant compound of Example 96A. The mixture was stirredfor 24 hours and then concentrated in vacuo. The residue was purified bypreparative thin layer chromatography using 10% methanol/CH₂Cl₂ aseluent followed by preparative reversed-phase C₁₈ HPLC using a lineargradient of 30% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to yield 50.2mg of the title compound. TLC: Rf=0.43, 10% methanol/CH₂Cl₂. HPLC:Rt=13.54 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 97

Compound 97. To solution of 60 mg of the resultant compound of Example35A in 1 mL of THF was added sequentially, 54 μL ofdiisopropylethylamine and a solution of 48.9 mg of the resultantcompound of Example 85A in 1 mL THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was diluted with 60 mLof CH₂Cl₂, washed with 5% sodium bicarbonate and brine, dried overmagnesium sulfate, filtered and concentrated in vacuo. The residue waspurified by preparative thin layer chromatography using 20% EtOAc/CH₂Cl₂as eluent to yield 46.9 mg of the title compound. TLC: Rf=0.31, 20%EtOAc/CH₂Cl₂. HPLC: Rt=15.18 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 98

Compound 98. To a solution of 61.0 mg of the resultant compound ofExample 35A in 1 mL of THF was added sequentially, 49 μL ofdiisopropylethylamine and a solution of 44 mg of the resultant compoundof Example 82A in 1 mL THF. The mixture was stirred for 24 hours andthen concentrated in vacuo. The residue was purified by preparative thinlayer chromatography using 5% methanol/CH₂Cl₂ as eluent to yield 61.0 mgof a white solid. TLC: Rf=0.19, 5 methanol/CH₂Cl₂. HPLC: Rt=13.28 min;13.28 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 99

Compound 99. A solution of 75 mg of the resultant compound of Example51D and 45 mg of 4-chlorobenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 24.6 mg of the title compoundwas obtained. TLC: Rf=0.3, 4% MeOH/CH₂Cl₂. HPLC: Rt=15.87 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 100

Compound 100. A solution of 40 mg of the resultant compound of Example51D and 45 mg of 4-methoxybenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 21.4 mg of the title compoundwas obtained as a white solid. TLC: Rf=0.2, 4% MeOH/CH₂Cl₂. HPLC:Rt=14.85 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 101

Compound 101. This compound was prepared from the resultant compound ofExample 128 by treatment with hydrogen chloride gas and subsequentreaction with the resultant compound of Example 48A in the mannerdescribed in Example 132. After workup and purification by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluant on a portion of the crude mixture, 4.2 mg of thetitle compound was obtained as a white solid. TLC: Rf=0.2, 4%MeOH/CH₂Cl₂. HPLC: Rt=11.53 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 102

Compound 102. A solution of 36 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of methyl chloroformate and 22 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 30% diethyl ether in CH₂Cl₂ aseluent to provide 27 mg of the title product as a white solid. TLC:Rf=0.10 (30% diethyl ether in CH₂Cl₂). HPLC: Rt=13.49 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 103

Compound 103. A solution of 29 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 6 mg of methyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 29 mg of the title product as a white solid. TLC:Rf=0.24 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.07 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 104

Compound 104. A solution of 31 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of methyl chloroformate and 21 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent is to provide 24 mg of the title product as a white solid. TLC:Rf=0.23 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=15.41 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 105

A. N-hydroxysuccinimidyl methallyl carbonate. To a solution of 2.9 mL of1.93 M phosgene in toluene at −10° C. was added 857 mg of methallylalcohol. The mixture was stirred for 2 h at −10° C. to produce a 1.9 Msolution of the title compound which was used directly in subsequentreactions.

B. Compound 105. A solution of 39 mg of the resultant compound ofExample 40A in CH₂Cl₂ was treated sequentially, at ambient temperatureunder an atmosphere of nitrogen, with 0.05 mL of the resultant compoundof Example 105A and 24 mg N,N-diisopropylethylamine. The mixture wasstirred for 3 h and then concentrated in vacuo. The residue was taken upin ethyl acetate and washed with 0.5 N HCl and saturated NaCl then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by preparative thin layer silica gel chromatography using ethylacetate as eluent to yield 18 mg of the title product as a white solid.TLC: Rf=0.67 (ethyl acetate). HPLC: Rt=14.97 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 106

Compound 106. A solution of 31 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 0.04 mL of the resultant compound ofExample 105A and 18 mg N,N-diisopropylethylamine. The mixture wasstirred for 3 h and then concentrated in vacuo. The residue was taken upin ethyl acetate and washed with 0.5 N HCl and saturated NaCl then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 5% diethylether/CH₂Cl₂ as eluent to provide 19 mg of the title product as a whitesolid. TLC: Rf=0.34 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=18.24 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 107

Compound 107. A solution of 28 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 0.05 mL of the resultant compound ofExample 105A and 19 mg N,N-diisopropylethylamine. The mixture wasstirred for 3 h and then concentrated in vacuo. The residue was taken upin ethyl acetate and washed with 0.5 N HCl and saturated NaCl then driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 5% diethylether in CH₂Cl₂ as eluent to provide 18 mg of the title product as awhite solid. TLC: Rf=0.25 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=16.68min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 108

Compound 108. To a solution of 62.5 mg of 124B in 1 mL of THF was addedsequentially 56 μL of diisopropylethylamine and a solution of 49.6 mg ofthe resultant compound of Example 82A in 1 mL THF. The mixture wasstirred for 24 hours and then concentrated in vacuo. The residue waspurified by preparative thin layer chromatography using 50% EtOAc/CH₂Cl₂as eluent followed by preparative reversed-phase C18 HPLC using a lineargradient of 30% to 100% CH₃CN/H₂O with 0.1% TFA as eluant on a portionof the crude mixture, 4.2 mg of the title compound was obtained as awhite solid. TLC: Rf=0.16, 10% methanol/CH₂ Cl₂. HPLC: Rt=13.67 min.(¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 109

A. (S)-4-Methoxycarbonyl-oxazlidin-2-one. To a solution of 4.88 g ofserine methyl ester hydrochloride in 25 mL of water was added 6.94 g ofpotassium carbonate. The mixture was cooled to 0° C. and 19.5 mL ofphosgene was added dropwise. After stirring at 0° C. for 3 hours, waterwas removed to yield a white solid with was washed with copious ofCH₂Cl₂. The organic solution was then dried over magnesium sulfate,filtered and concentrated to yield 3.26 g of the title product as aclear oil. (¹H) NMR (D₂O) δ=3.82 (s, 3H), 4.43 (dd, 1H), 4.53 (dd, 1 h),4.67 (t, 1H), 6.29 (s, 1H).

B. (S)-4-Hydroxymethyl-oxazolidin-2-one. To a solution of 3.26 g of theresultant compound of Example 109A in 20 mL of ethanol at 0° C. wasadded 0.85 g of sodium borohydride in small portions. The ice bath wasremoved and after additional 3 hours, 20 mL of 2.0 N hydrogen chloridewas added to the mixture, which was then concentrated to yield an oil.The residue was extracted with EtOAc and the organic solution was driedover magnesium sulfate, filtered and concentrated to yield 2.50 g of thetitle compound. (¹H) NMR (CDCl₃) δ=2.48 (s, 1H), 3.69 (dd, 1H), 4.08 (m,1H), 4.31 (t, 1H), 4.57 (t, 1H).

C. 4-Nitrophenyl-((S)-4-oxazolidin-2-onyl)-methyl carbonate. To asolution of 1.04 g of p-nitrophenyl chloroformate in 20 mL of CH₂Cl₂ at0° C. was added sequentially, 0.5 g of the resultant compound of Example109B and 0.6 mL of 4-methyl morpholine. The mixture was stirred for 2hours at ambient temperature and then concentrated in vacuo. The residuewas purified by low pressure silica gel column chromatography using 20%EtOAc in CH₂Cl₂ eluent to yield 0.57 g of the title compound. TLC:Rf=0.10, 50% EtOAc/hexane.

D. Compound 109. To a solution of 60 mg of the resultant compound ofExample 35A in 1 mL of THF was added sequentially, 56 μL ofdiisopropylethylamine and a solution of 51.1 mg of the resultantcompound of Example 109C in 1 mL acetonitrile. The mixture was stirredfor 24 hours and then concentrated in vacuo. The residue was purified bypreparative thin layer chromatography using 5% methanol/CH₂Cl₂ as eluentto yield 60.4 mg of the title compound. TLC: Rf=0.38, 5%methanol/CH₂Cl₂. HPLC: Rt=14.11 min. (¹H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 110

Compound 110. To a solution of 60 mg of the resultant compound ofExample 40A in 1 mL of acetonitrile was added sequentially, 51 μL ofdiisopropylethylamine and a solution of 46.8 mg of the resultantcompound of Example 109C in 1 mL acetonitrile. The mixture was stirredfor 48 hours and then concentrated in vacuo. The residue was purified bypreparative thin layer chromatography using 10% methanol/CH₂Cl₂ eluentfollowed by preparative reversed-phase C18 HPLC using a linear gradientof 30% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to yield 16 g of thetitle compound. TLC: Rf=0.28, 50% EtOAc/CH₂Cl₂. HPLC: Rt=12.47 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 111

A solution of 0.067 mmol of the resultant compound of Example 114D in 5mL of tetrahydrofuran was added 20 μL of diisopropylethylamine followeddropwise by a solution of the resultant compound of Example 82A in 5 mLof tetrahydrofuran during one hour. The mixture was stirred 16 h andthen concentrated in vacuo. The crude residue was purified by thicklayer silica gel chromatography using 5% MeOH/CH₂Cl₂ as eluant to obtain21.8 mg of the title compound. TLC: Rf=0.45, 5% MeOH/CH₂Cl₂; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 112

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=3-sulfonamidophenyl). To a solution of 96.6 mg (0.287 mmol) of theresultant compound of Example 39A in CH₂Cl₂ (4 mL) was added aqueoussodium bicarbonate (1 mL), solid sodium bicarbonate 36.2 mg (0.431mmol), and m-benzene disulfonylchloride 86.9 mg, (1.08 mmol). Afterstirring for 1 h, 30% ammonium hydroxide (10 mL) was added. After 14 hthe resulting mixture was diluted with CH₂Cl₂, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by flash chromatography using 0% to 10%methanol/CH₂Cl₂ eluent to provide 49.3 mg of the title product (¹H)-NMR(CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=isobutyl, E=3-sulfonamidophenyl, hydrochloridesalt). A solution of 49.3 mg (0.089 mmol) of the resultant compound ofExample 112A in EtOAc (10 mL) at −20° C. was treated with anhydrous HClgas for 10 min. The ice bath was removed and after an additional 15min., the reaction mixture was sparged with nitrogen then concentratedin vacuo to provide 53.1 mg of title product as the HCl salt. (¹H)-NMR(CDCl₃) consistent with structure.

C. Compound 112. To a solution of 53.1 mg of the resultant compound ofExample 112B (0.089 mmol) in CH₂ Cl₂ (3 mL) was treated sequentially atambient temperature under an atmosphere of nitrogen, with 0.031 mL(0.177 mmol) diisopropylethylamine and 24.3 mg (0.106 mmol) of theresultant compound of Example 48A. The mixture was stirred 16 h and thenconcentrated in vacuo. The residue was taken up in CH₂Cl₂ and washedwith saturated brine, dried over magnesium sulfate, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel column chromatography using a gradient 5% to 20% EtOAc in CH₂Cl₂ aseluent to yield 10.8 mg of the title product. TLC: RF=0.4 25% EtOAc inCH₂Cl₂. HPLC: Rt=13.3 min; (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 113

A. 3-Furansulfonyl chloride. In flame dried glassware under a nitrogenatmosphere to a solution of 428 mg (2.909 mmol) of 3 bromofuran inanhydrous tetrahydrofuran at −78° C. was added 2.0 mL n-butyl lithium(3.2 mmol at 1.6 molar in hexane). After 45 minutes the resultantsolution was added via cannula to a 20° C. solution of sulfuryl chloridein diethyl ether (5 mL plus a 2 mL rinse). After 1 h, the reaction wasquenched with 0.5 N hydrochloric acid and extracted into diethyl ether.The ethereal extracts were washed with saturated brine, dried overmagnesium sulfate, filtered and concentrated in vacuo to provide 158 mgof the title product. (¹H) NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl, E=3-furyl). To asolution of 289.7 mg (0.861 mmol) of the resultant compound of Example39A in CH₂Cl₂ (8 mL) was added aqueous sodium bicarbonate (2 mL), solidsodium bicarbonate 108 mg (1.292 mmol), and the resultant product fromExample 113A 157.8 mg, (1.08 mmol). After stirring for 1 h 30% ammoniumhydroxide (10 mL) was added. After 14 h, the resulting mixture wasdiluted with CH₂Cl₂, washed with saturated brine, dried over magnesiumsulfate, filtered and concentrated in vacuo. The residue was purified byflash chromatography using 1% to 15% EtOAc/CH₂Cl₂.

C. Compound XXII (A=H, D′=isobutyl, E=3-furyl, hydrochloride salt). Asolution of 217.3 mg (0.581 mmol) of the resultant compound of Example113B in EtOAc (15 mL) at −20° C. was treated with anhydrous HCl gas for10 min. The ice bath was removed and after an additional 15 min. thereaction mixture was sparged with nitrogen then concentrated in vacuoto, provide 228 mg of title product as the HCl salt. TLC: Rf=0.52 10%methanol/CH₂Cl₂.

D. Compound 113. To a solution of 65.3 mg of the resultant compound ofExample 113C (0.162 mmol) in CH₂Cl₂ (3 mL) was treated sequentially atambient temperature under an atmosphere of nitrogen, with 0.056 mL(0.324 mmol) diisopropylethylamine and 44.6 mg (0.194 mmol) of theresultant compound of Example 48A. The mixture was stirred 16 h and thenconcentrated in vacuo. The residue was taken up in CH₂Cl₂ and washedwith saturated brine, dried over magnesium sulfate, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel column chromatography using a gradient 3% to 20% EtOAc in CH₂Cl₂eluent to yield 10.8 mg of the title product. TLC: Rf=0.6, 25%EtOAc/CH₂Cl₂. HPLC: Rt=13.9 min; (¹H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 114

A. Aminomethylcyclopentane. To a solution of LiAlH₄ (38 g, 1.0 mole) indiethyl ether (2 L) was added cyclopentanecarbonitrile (73.2 g, 0.77mol) as a solution in 250 mL ether. The solution was stirred overnightat ambient temperature and then quenched by addition of the organics to3 L of a saturated potassium, sodium tartrate solution. The amine wasextracted into 3 L of ether, dried over anhydrous K₂CO₃ thenconcentrated by distillation to approximately 400 mL total volume. Thecrude product was purified via distillation to give 58.2 g of the titlecompound as a colorless oil. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXI (P=tert-butoxycarbonyl, D′=cyclopentylmethyl, P′═H). Tothe resultant compound of Example 114A (20 g, 0.2 mol) was addedcompound XX (P=Boc) (5.84 g) and the mixture was stirred for 24 h atambient temperature. The solution was concentrated by distillation underreduced pressure. The residue was triturated with hexane and the solidcollected by suction filtration and washed with hexane to give 7.08 g ofa white solid which was used without further purification. TLC: Rf=0.59(1:10:90 concentrated NH₄OH/methanol/CH₂Cl₂), (¹H)-NMR (CDCl₃)consistent with structure.

C. Compound XXII (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=4-fluorophenyl). To a solution of the resultant compound of Example114B (200 mg, 0.55 mmol) in CH₂Cl₂ (10 mL) was added4-fluorobenzenesulfonyl chloride (210 mg, 1.1 mmol) followed by theaddition of a saturated solution of sodium bicarbonate (3 mL) followedby addition of solid sodium bicarbonate (0.1 g, 1.2 mmol). The mixturewas allowed to stir at ambient temperature overnight. The solution wasdiluted with 100 mL CH₂Cl₂, the organics separated, dried over anhydrousMgSO₄, and the organics concentrated under reduced pressure to obtain0.33 g of crude product. This material was purified via medium pressureliquid chromatography using CH₂Cl₂, followed by 0.5:99.5methanol/CH₂Cl₂, followed by a 1:99 methanol/CH₂Cl₂ solution as thesolvent system to give 120 mg (42% yield) of the title compound as awhite solid. TLC: Rf=0.48 (3:97 methanol/CH₂Cl₂); HPLC: Rt=18.22 min,(¹H)-NMR (CDCl₃) consistent with structure.

D. Compound XXII (P═H, D′=cyclopentylmethyl, E=4-fluorophenyl,hydrochloride salt). A solution of 266 mg of the resultant compound ofExample 114C in ethyl acetate was treated at −20° C. with HCl gas for 20min, during which time the temperature was allowed to warm to 20° C.Nitrogen was then bubbled through the mixture for 15 min and the solventremoved in vacuo to yield 224 mg of white solid which was used directlyfor ensuing reaction.

E. Compound 114. A solution of 31 mg of the resultant compound ofExample 114D in CH₂Cl₂ was treated sequentially, at ambient temperature,under an atmosphere of nitrogen, with 9 mg of allyl chloroformate and 19mg N,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed in 0.5 N HCl and saturated NaCl then dried over MgSO₄, filtered,and concentrated in vacuo to yield 34 mg of the title product as a whitesolid. TLC: Rf=0.34 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.21 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 115

Compound 115. A solution of 31 mg of the resultant compound of Example114B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of ethyl chloroformate and 19 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo to yield 35 mg of the title productas a white solid. TLC: Rf=0.32 (5% diethyl ether/CH₂Cl₂). HPLC: Rt=16.86min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 116

A. Compound XXII (A=tert-butoxycarbonyl, D′cyclopentylmethyl,E=4-chlorophenyl). The resultant compound of Example 114B (252 mg) wasreacted with 4-chlorobenzenesulfonyl chloride (175 mg) in the mannerdescribed in Example 166A. Workup and purification by silica gelchromatography using EtOAc/CH₂Cl₂ as eluant yielded the product as awhite solid; (1H) NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=cyclopentylmethyl, E=4-chlorophenyl,hydrochloride salt). A solution of 320 mg of the resultant compound ofExample 116A in 20 mL of EtOAc was treated with anhydrous HCl gas for 5min. The reaction mixture was sparged with nitrogen then concentrated invacuo to yield a white solid which was used directly for subsequentreaction.

C. Compound 116. To a solution of 63.4 mg of the resultant compound ofExample 116B in 1 mL of THF was added sequentially 54 μL ofdiisopropylethylamine and a solution of 39.9 mg of the resultantcompound of Example 48A in 1 mL THF. The mixture was stirred for 24hours and then concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 20% EtOAc in CH₂Cl₂eluent to yield 0.62 g of the title compound. TLC: Rf=0.71, 40%EtOAc/CH₂Cl₂. HPLC: Rt=16.88 min. (¹H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 117

Compound 117. A solution of 66.1 mg of the resultant compound of Example116B in 1 mL of THF was treated sequentially with 56 μL ofdiisopropylethylamine and 19.3 μL of allyl chloroformate. The mixturewas stirred for 4 hours and concentrated in vacuo. The residue was takeninto 50 mL of EtOAc and washed with 1.0 N HCl, saturated sodiumbicarbonate, brine, dried over magnesium sulfate, filtered andconcentrated. The residue was purified by low pressure silica gel columnchromatography using 20% EtOAc in hexane eluent to yield 69.7 mg of thetitle compound. TLC: Rf=0.20, 20% EtOAc/hexane. HPLC: Rt=17.83 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 118

Compound 118. To a solution of 65.3 mg of the resultant compound ofExample 116B in 1 mL of THF was added sequentially 55 μL ofdiisopropylethylamine and a solution of 49.2 mg of the resultantcompound of Example 82A in 1 mL THF. The mixture was stirred for 24hours and concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 40% EtOAc in CH₂Cl₂ aseluent followed by preparative reversed-phase C₁₈ HPLC using a lineargradient of 40% to 80% CH₃CN/H₂O for elution to yield 70.7 mg of thetitle compound. TLC: Rf=0.27, 40% EtOAc/CH₂Cl₂. HPLC: Rt=14.85 min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 119

Compound 119. A solution of 26 mg of the resultant compound of Example81B in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 6 mg of ethyl chloroformate and 15 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetage andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 26 mg of the title product as a white solid. TLC:Rf=0.19 (51 diethyl ether in CH₂Cl₂). HPLC: Rt=17.50 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 120

Compound 120. A solution of 30 mg of the resultant compound of Example40A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of ethyl chloroformate and 18 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetage andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified bypreparative thin layer silica gel chromatography using ethyl acetate aseluent to yield 25 mg of the title product as a white solid. TLC:Rf=0.60 (ethyl acetate). HPLC: Rt=13.86 min. (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 121

Compound 121. A solution of 26 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 7 mg of ethyl chloroformate and 17 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 22 mg of the title product as a white solid. TLC:Rf=0.14 (5% diethyl ether/CH₂Cl₂). HPLC Rt=15.95 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 122

Compound 122. A solution of 27 mg of the resultant compound of Example35A in CH₂Cl₂ was treated sequentially, at ambient temperature under anatmosphere of nitrogen, with 8 mg of allyl chloroformate and 18 mgN,N-diisopropylethylamine. The mixture was stirred for 3 h and thenconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with 0.5 N HCl and saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether in CH₂Cl₂ aseluent to provide 23 mg of the title product as a white solid. TLC:Rf=0.33, 5% diethyl ether in CH₂Cl₂. HPLC: Rt=16.28 min. (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 123

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=3,4-dimethoxyphenyl). To a solution of 401 mg (1.192 mmol) of theresultant compound of Example 39A in CH₂Cl₂ (12 mL) was added aqueoussodium bicarbonate (3 mL), solid sodium bicarbonate 130 mg (1.549 mmol),and 3,4-dimethoxybenzenesulfonyl chloride 33.8 mg, (1.43 mmol). After 14h, the resulting mixture was diluted with EtOAc, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by flash chromatography using 5% to 25%EtOAc/CH₂Cl₂ eluent to provide 440.1 mg of the title product. TLC:Rf=0.72, 20% EtOAc/CH₂Cl₂.

B. Compound XXII (A=H, D′=isobutyl, E=3,4-dimethoxyphenyl, hydrochloridesalt). A solution of 440 mg (0.820 mmol) of the resultant compound ofExample 123A in EtOAc (15 mL) at −20° C. was treated with anhydrous HClgas for 10 min. The ice bath was removed and after an additional 15 min.the reaction mixture was sparged with nitrogen then concentrated invacuo to provide 610 mg of title product as the HCl salt. TLC: Rf=0.44,10% methanol/CH₂Cl₂.

C. Compound 123. A solution of 38.9 mg of the resultant compound ofExample 123B (0.170 mmol) in CH₂Cl₂ (3 mL) was treated sequentially atambient temperature under an atmosphere of nitrogen with 0.049 mL (0.283mmol) diisopropylethylamine and 66.9 mg (169.6 mmol) of the resultantcompound of Example 48A. The mixture was stirred for 16 h and thenconcentrated in vacuo. The residue was taken up in CH₂Cl₂ and washedwith saturated brine, dried over magnesium sulfate, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel column chromatography using a gradient 10% to 25% diethyl ether inCH₂Cl₂ eluent to yield 57.6 mg of the title product. TLC: Rf=0.39, 25%diethyl ether/CH₂Cl₂. HPLC: Rt=14.3 min; (1H) NMR (CDCl₃) consistentwith structure.

EXAMPLE 124

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl, E=3,4difluorophenyl). To a solution of 332.7 mg (0.989 mmol) of the resultantcompound of Example 39A in CH₂Cl₂ (12 mL) was added aqueous sodiumbicarbonate (3 mL), solid sodium bicarbonate 125 mg (1.483 mmol), and3,4 difluorobenzensulfonyl chloride 231 mg. (1.088 mmol). After 14 h,the resulting mixture was diluted with CH₂Cl₂, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by flash chromatography using 5% to 25% diethylether/CH₂Cl₂ eluent to provide 313.6 mg of the title product. (¹H)-NMR(CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=isobutyl, E=3,4 difluorophenyl, hydrochloridesalt). A solution of 312.6 mg (0.610 mmol) of the resultant compound ofExample 124A in EtOAc (15 mL) at −20° C. was treated with anhydrous HClgas for 10 min. The ice bath was removed and after an additional 15min., the reaction mixture was sparged with nitrogen then concentratedin vacuo to provide 280 mg of title product as a white solid. TLC:Rf=0.46, 10% methanol/CH₂Cl₂.

C. Compound 124. To a solution of 64.7 mg of the resultant compound ofExample 124B (0.144 mmol) in CH₂Cl₂ (3 mL) was treated sequentially atambient temperature under an atmosphere of nitrogen, with 0.050 mL(0.288 mmol) diisopropylethylamine and 39.6 mg (172.9 mmol) of theresultant compound of Example 48A. The mixture was stirred for 16 h andthen concentrated in vacuo. The residue was taken up in CH₂Cl₂ andwashed with saturated brine, dried over magnesium sulfate, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel column chromatography using a gradient 5% to 20% diethyl ether inCH₂Cl₂ eluent to yield 44 mg of the title product. TLC: RF=0.54 25%diethyl ether/CH₂Cl₂. HPLC: Rt=15.4 min. (H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 125

Compound 125. This compound was prepared from the resultant compound ofExample 146B in the manner described in Example 88. After workup andpurification by preparative reversed-phase C₁₈ HPLC using a linergradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant, 10.5 mg ofthe title compound was obtained as a white solid. TLC: Rf=0.4, 4%MeOH/CH₂Cl₂. HPLC: Rt=14.06 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 126

A. Compound XXI (P=tert-butoxycarbonyl, D′=methyl, P′═H). To a solutioncompound XX (1.7 mmol) in ethanol (20 mL) was added methylamine gas, atambient temperature, for 30 min. The solution was stirred overnight,then concentrated under reduced pressure to give 0.47 g of the titlecompound which was used without subsequent purification. TLC: Rf=0.19,1:10:90 NH₄OH/methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistent withstructure.

B. Compound 126. To a solution of the product of Example 126A (0.15 g,0.51 mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by addition of solid sodium bicarbonate (90mg, 1.1 mmol), followed by addition of 3,4-dichlorobenzenesulfonylchloride (0.25 g, 1.0 mmol). The mixture was stirred at ambienttemperature overnight. The organics were extracted into 100 mL CH₂Cl₂,dried over anhydrous, MgSO₄, concentrated under reduced pressure thenpurified via medium pressure silica gel chromatography using a gradientsystem of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂. The title compound wasobtained as a colorless foam 210 mg. TLC: Rf=0.42 (3:97methanol/CH₂Cl₂), HPLC: Rt=17.2 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 127

Compound 127. To a solution of the product of Example 126A (0.15 g, 0.51mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by addition of solid sodium bicarbonate(100 mg, 1.0 mmol), followed by addition of 4-fluorobenzenesulfonylchloride (0.20 g, 1.0 mmol). The mixture was stirred at ambienttemperature overnight. The organics were extracted into 100 mL CH₂Cl₂,dried over anhydrous, MgSO₄, concentrated under reduced pressure thenpurified via medium pressure silica gel chromatography using a gradientsystem of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂. The title compound wasobtained as a white solid 104 mg. TLC: Rf=0.36, 3.97 methanol/CH₂Cl₂,HPLC: Rt=15.86 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 128

Compound 128. To a solution of the product of Example 126A (0.15 g, 0.51mmol) in CH₂Cl₂ (6 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by addition of solid sodium bicarbonate (90mg, 1.0 mmol), followed by addition of acetamidobenzenesulfonyl chloride(0.24 g, 1.02 mmol). The mixture was stirred at ambient temperatureovernight. The organics were extracted into 100 mL CH₂Cl₂, dried overanhydrous, MgSO₄, concentrated under reduced pressure then purified viamedium pressure silica gel chromatography using a gradient system ofCH₂Cl₂ followed by 5:95 EtOAc/CH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂.The title compound was obtained as 244 mg of white solid. TLC: Rf=0.13,3:97 methanol/CH₂Cl₂, HPLC: Rt=13.47 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 129

A. Compound XXI (P=tert-butoxycarbonyl, D′=(2-tetrahydrofuryl)-methyl,P′═H). To a solution compound XX (3.3 mmol) in ethanol (30 mL) was addedtetrahydrofurfurylamine (1.03 mL, 10 mmol). The mixture was warmed to85° C. and stirred overnight. The solution was filtered and the solutionconcentrated under reduced pressure to give 1.29 g of the title compoundwhich was used without subsequent purification. TLC: Rf=0.52, 1:10:90NH₄OH/methanol/CH₂Cl₂

B. Compound 129. To a solution of the resultant compound of Example 129A(200 mg, 0.55 mmol) in CH₂Cl₂ (6 mL) was added 4-fluorobenzenesulfonylchloride (320 mg, 1.6 mmol) followed a saturated solution of sodiumbicarbonate (3 mL) and solid sodium bicarbonate (0.1 g, 1.2 mmol). Themixture was stirred at ambient temperature overnight. The solution wasdiluted with 100 mL CH₂Cl₂, the organics separated, dried over anhydrousMgSO₄, and the organics concentrated under reduced pressure. The crudeproduct was purified via medium pressure liquid chromatography using agradient solvent system of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂ followedby a 10:90 ether/CH₂Cl₂ solution to give 130 mg of the title compound asa white solid. TLC: Rf=0.35, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.37 min,(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 130

A. Compound XXI (P=tert-butoxycarbonyl, D′=(isobutenyl, P′═H)). To asolution compound XX (P=tert-butoxycarbonyl)(2.5 mmol) in ethanol (30mL) was added a solution 2-methylallylamine hydrochloride (1.34 g, 12.5mmol) and KOH (0.70 g, 12.5 mmol) in ethanol (20 mL). The mixturestirred 30 min at ambient temperature. The solutions were combined andheated to 85° C. for 24 h. The solution was filtered and concentratedunder reduced pressure to give 0.82 g of the title compound which wasused without subsequent purification. TLC: Rf=0.45, 1:10:90 concentratedNH₄OH/methanol/CH₂ Cl₂.

B. Compound 130. To a solution of the product of Example 130A (0.20 g,0.60 mmol) in CH₂Cl₂ (6 mL) was added a saturated solution of sodiumbicarbonate (3 mL), followed by solid sodium bicarbonate (0.1 g, 1.2mmol) and then p-fluorobenzenesulfonyl chloride (0.35 g, 1.78 mmol). Themixture was stirred at ambient temperature for 24 h. The organics wereextracted into 100 mL CH₂C₂, dried over anhydrous MgSO₄, concentratedunder reduced pressure then purified via medium pressure silica gelchromatography using a gradient system of CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂. The title compound was obtained as a white solid 180mg. TLC: Rf=0.35, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.82 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 131

Compound 131. To a solution of the resultant compound of Example 130A(200 mg, 0.60 mmol) in CH₂Cl₂ (6 mL) was added4-acetamidobenzenesulfonyl chloride (410 mg, 1.76 mmol), followed by asaturated solution of sodium bicarbonate (3 mL) and solid sodiumbicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambienttemperature overnight. The solution was diluted with 100 mL CH₂Cl₂, theorganics separated, dried over anhydrous MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using a gradient solvent system ofCH₂Cl₂, followed by 30:70 EtOAc/CH₂Cl₂ solution to give 140 mg of thetitle compound as a white solid. TLC: Rf=0.19, 3:97 methanol/CH₂Cl₂,HPLC: Rt=15.06 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 132

A. Compound XXII (A=H, D′=(2-tetrahydrofuryl)-methyl, E=4-fluorophenyl,hydrochloride salt). To a solution of the resultant compound of Example129B (30 mg, 0.057 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc(1 mL). The mixture was stirred overnight at ambient temperature. Thesolution was concentrated under reduced pressure to give 16 mg of thetitle compound as a white solid which was used without subsequentpurification. TLC: Rf=0.60 (1:10:90 NH₄OH/methanol/CH₂Cl₂).

B. Compound 132. To a solution of the resultant compound of Example 132A(16 mg) in CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol)followed by the compound of Example 48A (20 mg, 0.09 mmol). The mixturewas stirred at ambient temperature for 24 hours. The solution wasconcentrated under reduced pressure and the crude product purified viamedium pressure column chromatography using 20:80 EtOAc/CH₂Cl₂ as thesolvent system to give 7.4 mg. Rf=0.37 (3:97 methanol/CH₂Cl₂), HPLC:Rt=14.19 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 133

A. Compound XXII (A=tert-butoxycarbonyl, D′ (2-tetrahydrofuryl)-methyl,E=4-acetamidophenyl). To a solution of the resultant compound of Example129A (200 mg, 0.55 mmol) in CH₂Cl₂ (6 mL) was added4-acetamidobenzenesulfonyl chloride (380 mg, 1.6 mmol) followed by asaturated solution of sodium bicarbonate (3 mL) and solid sodiumbicarbonate (0.1 g, 1.2 mmol). the mixture was stirred at ambienttemperature overnight. The solution was diluted with 100 mL CH₂Cl₂, theorganics separated, dried over anhydrous. MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using a gradient solvent system ofCH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂, followed by a 30:70 EtOAc/CH₂Cl₂solution to give 120 mg of the title compound as a white solid. TLC:Rf=0.13, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistent withstructure.

B. Compound XXII (A=H, D′=(2-tetrahydrofuryl)-methyl,E=4-acetamidophenyl, hydrochloride salt). To a solution of the resultantcompound of Example 133A (120 mg 0.22 mmol) in EtOAc (5 mL) was added30% w/w HCl in EtOAc (2 mL). The mixture was stirred overnight atambient temperature. The solution was concentrated under reducedpressure to give the title compound which was used without subsequentpurification. TLC: Rf=0.50, 1:10:90 NH₄OH/methanol/CH₂Cl₂.

C. Compound 133. To a solution of the resultant compound of Example 133Bin CH₂Cl₂ (5 mL) was added triethylamine (0.2 mL, 1.4 mmol) followed bythe compound of Example 48A (73 mg, 0.32 mmol). The mixture was stirredat ambient temperature for 24 hours. The solution was concentrated underreduced pressure and the crude product purified via medium pressurecolumn chromatography using a gradient solvent system of CH₂Cl₂,followed by 1:99 methanol/CH₂Cl₂, followed by 3:97 methanol/CH₂Cl₂ asthe solvent system to give 87.8 mg. Rf=0.09, 3:97 methanol/CH₂Cl₂, HPLC:Rt=12.53 min (₁H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 134

A. Compound XXII (A=H, D′=isobutenyl, E=4-acetamidophenyl, hydrochloridesalt). To a solution of the resultant compound of Example 131 (40 mg0.075 mmol) in EtOAc (5 mL) was added 30% w/w HCl in EtOAc (2 mL). Themixture was stirred overnight at ambient temperature. The solution wasconcentrated under reduced pressure to give the title compound, whichwas used without subsequent purification. TLC: Rf=0.38, 1:10:90NH₄OH/methanol/CH₂Cl₂.

B. Compound 134. To a solution of the resultant compound of Example 134Ain CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol), followedby the compound of Example 48A (26 mg, 0.11 mmol). The mixture wasstirred at ambient temperature for 24 hours. The solution wasconcentrated under reduced pressure and the crude product purified viamedium pressure column chromatography using a gradient solvent system ofCH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂, followed by 3:97methanol/CH₂Cl₂ as the solvent system to give 10.1 mg of the titlecompound. Rf=0.11 (3:97 methanol/CH₂Cl₂), HPLC: Rt=12.86 min, (₁H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 135

A. Compound XXI (A=H, D′=(isobutenyl, E=4-fluorophenyl, hydrochloridesalt). To a solution of the resultant compound of Example 130B (50 mg,0.10 mmol) in EtOAc (5 mL) was added 30% w/w HCl in EtOAc (1 mL). Themixture was stirred overnight at ambient temperature. The solution wasconcentrated under reduced pressure to give the title compound which wasused without subsequent purification. TLC: Rf=0.48, 1:10:90NH₄OH/methanol/CH₂Cl₂.

B. Compound 135. To a solution of the resultant compound of Example 135Ain CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol), followedby the compound of Example 48A (35 mg, 0.15 mmol). The mixture wasstirred at ambient temperature for 24 hours. The solution wasconcentrated under reduced pressure and the crude product purified viamedium pressure column chromatography using a gradient solvent system ofCH₂Cl₂ followed by 20:80 EtOAc/CH₂Cl₂ as the solvent system to give 12mg. Rf=0.34, 3:97 methanol/CH₂Cl₂, HPLC: Rt=14.64 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 136

A. Compound XXI (A=tert-butoxycarbonyl, D′=2-furfuryl, A′=H). To asolution compound XX (2.5 mmol) in ethanol (30 mL) was addedfurfurylamine (0.67 mL, 7.5 mmol) and the mixture was heated to 85° C.for 24 h. The solution was filtered and concentrated under reducedpressure to give 0.80 g of the title compound which was used withoutsubsequent purification. TLC: Rf=0.38, 1:10:90 concentratedNH₄OH/methanol/CH₂Cl₂.

B. Compound XXII (A=tert-butoxycarbonyl, D′=2-furyl, E=4-fluorophenyl).To a solution of the product of Example 136A (0.20 g, 0.60 mmol) inCH₂Cl₂ (6 mL) was added a saturated solution of sodium bicarbonate (3mL), followed by addition of solid sodium bicarbonate (0.1 g, 1.2 mmol),then p-fluorobenzenesulfonyl chloride (0.32 g, 1.6 mmol). The mixturewas stirred at ambient temperature for 24 h. The organics were extractedinto 100 mL CH₂Cl₂, dried over anhydrous MgSO₄, concentrated underreduced pressure, then purified via medium pressure silica gelchromatography using a gradient system of CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂. The title compound was obtained as a white solid (86.1mg). TLC: Rf=0.17, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.5 min; (₁H)-NMR(CDCl₃) consistent with structure.

C. Compound XXII (A=H, D′=2-furyl, E=4-fluorophenyl, hydrochloridesalt). To a solution of the resultant compound of Example 136B (16 mg,0.031 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc (1 mL). Themixture was stirred overnight at ambient temperature. The solution wasconcentrated under reduced pressure to give the title compound, whichwas used without subsequent purification. TLC: Rf=0.48, 1:10:90NH₄OH/methanol/CH₂Cl₂.

D. Compound 136. To a solution of the resultant compound of Example 136Cin CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol), followedby the resultant compound of Example 48A (11 mg, 0.05 mmol). The mixturewas stirred at ambient temperature for 24 hours. The solution wasconcentrated under reduced pressure and the crude product purified viamedium pressure column chromatography using a gradient solvent system ofCH₂Cl₂ followed by 20:80 EtOAc/CH₂Cl₂ as the solvent system to give 4.9mg. TLC: Rf=0.28, (3:97 methanol/CH₂Cl₂, HPLC: Rt=14.57 min, (₁H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 137

A. Compound XXII (A=tert-butoxycarbonyl, D′=2-furyl,E=4-acetamidophenyl). To a solution of the resultant compound of Example136B (200 mg, 0.55 mmol) in CH₂Cl₂ (6 mL) was added4-acetamidobenzenesulfonyl chloride (390 mg, 1.7 mmol) followed bysaturated solution of sodium bicarbonate (3 mL) and solid sodiumbicarbonate (0.1 g, 1.2 mmol). The mixture was stirred at ambienttemperature overnight. The solution was diluted with 100 mL CH₂Cl₂, theorganics separated, dried over anhydrous. MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using a gradient solvent system ofCH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂, followed by a 30:70 EtOAc/CH₂Cl₂solution to give 100 mg of the title compound as a white solid. TLC:Rf=0.19, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistent withstructure.

B. Compound XXII (A=H, D′=2-furyl, E=4-acetamidophenyl, hydrochloridesalt). To a solution of the resultant compound of Example 137A (30 mg,0.054 mmol) in EtOAc (3 mL) was added 30% w/w HCl in EtOAc (1 mL). Themixture was stirred overnight at ambient temperature. The solution wasconcentrated under reduced pressure to give the title compound which wasused without subsequent purification. TLC: Rf=0.37 (1:10:90 NHOH/methanol/CH₂Cl₂.

C. Compound 137. To a solution of the resultant compound of Example 137ain CH₂Cl₂ (5 mL) was added triethylamine (0.1 mL, 0.72 mmol) followed bythe compound of Example 48A (19 mg, 0.083 mmol). The mixture was stirredat ambient temperature for 24 hours. The solution was concentrated underreduced pressure and the crude product purified via medium pressurecolumn chromatography using a gradient solvent system of CH₂Cl₂ followedby 1:99 methanol/CH₂Cl₂, followed by 3:97 methanol/CH₂Cl₂ as the solventsystem to give 8.5 mg of the title compound. TLC: Rf=0.11 (3:97methanol/CH₂Cl₂), HPLC: Rt=12.69 min; (₁H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 138

Compound 138. A solution of 75 mg of the resultant compound of Example51D and 45 mg of 3-chlorobenzenesulfonyl chloride were reacted in themanner described in Example 60. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 29.7 mg of the title compoundwas obtained. TLC: Rf=0.3, 4% MeOH/CH₂Cl₂, HPLC: Rt=15.83 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 139

Compound 139. To a solution of 67.9 mg of the resultant compound ofExample 116B in 1 mL of THF was added sequentially, 57 μL ofdiisopropylethylamine and a solution of 52.6 mg of the resultantcompound of Example 109C in 1 mL THF. The mixture was stirred for 24hours and concentrated in vacuo. The residue was purified by preparativethick layer silica gel chromatography using 7% methanol in CH₂Cl₂ eluentto yield 70.0 mg of the title compound. TLC: Rf=0.30, 5%methanol/CH₂Cl₂. HPLC: Rt=15.78 min; (¹H) NMR (CDCl₃) consistent withstructure.

EXAMPLE 140

A. 3(S)-amino-2(syn)-hydroxy-4-phenyl-1-chlorobutane formate salt. To aslurry of 16.33 g of 10% palladium on carbon (25% by weight) in methanoland tetrahydrofuran (400 mL, 1:1) was added, under N₂, 65.35 g of3(S)-N-(-benzyloxycarbonyl)-amino-1-chloro-2(syn)-hydroxy-4-phenylbutane(195.77 mmol) as a solution in methanol and tetrahydrofuran (1.2 L). Tothis slurry was added 540 mL of formic acid. After 15 h, the reactionmixture was filtered through diatomaceous earth and concentrated todryness. The resultant oil was slurried in toluene and evaporated, thentriturated sequentially with diethyl ether and CH₂Cl₂ to provide 47.64 gof product as a granular tan solid. TLC: Rf=0.17, 5% acetic acid/ethylacetate.

B.3(S)-N-(3(S)-tetrahydrofuryloxycarbonyl)-amino-1-chloro-2(syn)-hydroxy-4-phenylbutane.To a solution of the resultant compound of Example 140A (1.97 g, 7.95mmol) in CH₂Cl₂ (20 mL) was added a saturated solution of sodiumbicarbonate (5 mL), followed by solid sodium bicarbonate (1.33 g, 17.9mmol), and the resultant compound of Example 48A (2.0 g, 8.7 mmol). Themixture was stirred at ambient temperature overnight. The solution wasdiluted with 200 mL CH₂Cl₂, the organics separated, dried over anhydrousMgSO₄, and concentrated under reduced pressure. The residue wasrecrystallized from ethyl acetate/hexane to give 1.01 g of the titlecompound as a white solid. TLC: Rf=0.35, 3:97 methanol/CH₂Cl₂. (¹H)-NMR(CDCl₃) consistent with structure.

C. Compound XX (A=3(s)-tetrahydrofuryloxycarbonyl). To a solution of theresultant compound of Example 140B (1.0 g, 3.2 mmol) in absolute ethanol(15 mL) was added solid KOH (0.21 g, 3.8 mmol). The mixture was stirredat ambient temperature for 1.0 h. The solution was filtered through apad of Celite then concentrated under reduced pressure. The residue wastaken up in ether (100 mL), washed with brine, dried over MgSO₄, theconcentrated under reduced pressure to give 0.88 g of the title compoundas a white solid. TLC: Rf=0.49 (3:97 methanol/CH₂Cl₂), (¹H)-NMR (CDCl₃)consistent with structure.

D. Compound XXI (A (S)-3-tetrahydrofuryloxycarbonyl,D′=cyclopentylmethyl, A′=H). The resultant compound of Example 140C(0.88 g, 3.2 mol) was added to the resultant compound of Example 114A(5.0 g, 50.4 mmol) and stirred for 24 h at ambient temperature. Thesolution was concentrated by distillation under reduced pressure. Theresidue was triturated with hexane and the solid collected by suctionfiltration and washed with hexane to give 0.93 g of the title compound.TLC: Rf=0.44, 1:10:90 concentrated NH₄OH/methanol/CH₂Cl₂; (¹H)-NMR(CDCl₃) consistent with structure.

E. Compound 140. To a solution of the resultant compound of Example 140D(0.93 g, 2.47 mmol) in CH₂Cl₂ (20 mL) was added a saturated solution ofsodium bicarbonate (5 mL) followed by addition of solid sodiumbicarbonate (0.42 g, 4.94 mmol) and 4-methoxybenzenesulfonyl chloride(0.61 g, 2.96 mmol). the mixture was stirred at ambient temperature for4 hours. The solution was diluted with 200 mL CH₂Cl₂, the organicsseparated, dried over anhydrous MgSO₄, and the organics concentratedunder reduced pressure. The crude product was purified via mediumpressure liquid chromatography using CH₂Cl₂ followed by 1:99methanol/CH₂Cl₂ solution as the eluent system to give 1.28 g of thetitle compound as a white solid. TLC: Rf=0.26, 3:97 methanol/CH₂Cl₂,HPLC: Rt=15.66 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 141

A. Compound XXII (A=H, D′=cyclopentylmethyl, E=4-methoxyphenyl,hydrochloride salt). A solution of 71.3 mg of the resultant compound ofExample 166A in EtOAc (25 mL) at 0° C. was treated with anhydrous HClgas for 10 min., and allowed to stand for 12 h while warming to ambienttemperature, then concentrated under reduced pressure and the resultingwhite solid used without purification for subsequent reaction.

B. Compound 141. The resultant compound of Example 141A (0.134 mmol) wasreacted with allyl chloroformate in the manner described in Example 87B.After concentration of the mixture in vacuo and workup, the residue waspurified by thick layer silica gel chromatography using 5% MeOH/CH₂Cl₂as eluant followed by preparative reversed-phase C₁. HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 21.6mg of the title compound. TLC: Rf=0.45, 5% MeOH/CH₂Cl₂. HPLC: Rt=16.96min.

EXAMPLE 142

Compound 142. To a solution of 4.0 g of the resultant compound ofExample 141A in 45 mL of THF was added sequentially, 1.96 mL ofdiisopropylethylamine and a solution of 2.68 g of the resultant compoundof Example 82A in 45 mL THF. The mixture was stirred for 24 hours andconcentrated in vacuo. The residue was taken up in CH₂Cl₂, washed withsaturated sodium bicarbonate and brine, dried over magnesium sulfate,filtered and concentrated in vacuo. The residue was purified by lowpressure silica gel column chromatography using 20% to 40% EtOAc inhexane eluent to yield 3.69 g of the title compound. TLC: Rf=0.41, 50%EtOAc/CH₂Cl₂.

EXAMPLE 143

Compound 143. A solution of 3.69 g of the resultant compound of Example142 in 100 mL of ethyl ether was treated with anhydrous HCl gas for 10min. The reaction mixture was sparged with nitrogen then filtered. Thesolid was taken up in methanol and concentrated to yield 3.71 g of thetitle compound. TLC: Rf=0.62, 90/10/1 CH₂Cl₂/MeOH/AcOH, HPLC: Rt=13.87min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 145

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=2-(5-isoxazoy-3-yl)-thiophene). To a solution of 342.5 mg (1.02 mmol)of the resultant compound of Example 39A in CH₂Cl₂ (8 mL) was addedaqueous sodium bicarbonate (2 mL), solid sodium bicarbonate 257 mg (3.1mmol), and 5-(isoxazol-3-yl)-thiophenesulfonyl chloride 254.2 mg, (1.02mmol). After 14 h, the resulting mixture was diluted with CH₂Cl₂, washedwith saturated brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by flash chromatographyusing 5% to 25% EtOAc/CH₂Cl₂ eluent and recrystallized from ether CH₂Cl₂to provide 228.6 mg of the title product. (¹H)-NMR (CDCl₃) consistentwith structure.

B. Compound XXII (A-H, D′=isobutyl, E=2-(5-isoxazoy-3-1y)-thiopene,hydrochloride salt). A solution of 228.6 mg (0.416 mmol) of theresultant compound of Example 145A in EtOAc (15 mL) at −20° C. wastreated with anhydrous HCl gas for 10 min. The ice bath was removed andafter an additional 15 min, the reaction mixture was sparged withnitrogen then concentrated in vacuo to provide 223.6 mg of title productas the HCl salt. TLC: Rf=0.48, 10% methanol/CH₂Cl₂.

C. Compound 145. A solution of 78.5 mg of the resultant compound ofExample 145B (0.162 mmol) in CH₂Cl₂ (3 mL) was treated sequentially atambient temperature under an atmosphere of nitrogen with 0.07 mL (0.408mmol) diisopropylethylamine and 55.6 mg (0.243 mmol) of the resultantcompound of Example 48A. The mixture was stirred 16 h and thenconcentrated in vacuo. The residue was taken up in CH₂Cl₂ and washedwith saturated brine, dried over magnesium sulfate, filtered, andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 48.7 mg of the title product. TLC: Rf=0.36, 25% EtAOc/CH₂Cl₂.HPLC: Rt=15.2 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 146

A. Compound XXI (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=4-acetamidophenyl). To a solution of the resultant compound of Example114B (300 mg, 0.83 mmol) in CH₂Cl₂ (15 mL) was added4-acetamidobenzenesulfonyl chloride (580 mg, 2.48 mmol) followed by theaddition of a saturated solution of sodium bicarbonate (4 mL) and solidsodium bicarbonate (0.14 g, 1.67 mmol). The mixture was stirred atambient temperature overnight. The solution was diluted with 150 mLCH₂Cl₂, the organics separated, dried over anhydrous. MgSO₄, and theorganics concentrated under reduced pressure. The crude product waspurified via medium pressure liquid chromatography using a gradientsolvent system of CH₂Cl₂, followed by 5:95 EtOAc/CH₂Cl₂, followed by10:90 EtOAc/CH₂Cl₂ solution to give 310 mg of the title compound as awhite solid. TLC: Rf=0.10, 3:97 methanol/CH₂Cl₂, HPLC: Rt=15.96 min,(¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (P═H, D′=cyclopentylmethyl, E=4-acetamidophenyl,hydrochloride salt). To a solution of the resultant compound of Example146A (210 mg, 0.38 mmol) was added 30% w/w HCl in EtOAc (15 mL). Themixture was stirred for 1 hour at ambient temperature. The solution wasconcentrated under reduced pressure to give 180 mg of the title compoundwhich was used without subsequent purification. TLC: Rf=0.14, 1:10:90NH₄OH/methanol/CH₂Cl₂.

C. Compound XXII (P=allyloxycarbonyl, D′=cyclopentylmethyl,E=4-acetamidophenyl). To a solution of the resultant compound of Example146B (100 mg, 0.20 mmol) in CH₂Cl₂ (10 mL) was added triethylamine (0.1mL, 0.72 mmol), followed by allylchloroformate (0.04 mL, 0.3 mmol). Themixture was stirred at ambient temperature for 24 hours. The solutionwas diluted with 150 mL CH₂Cl₂, washed with water, dried over anhydrousMgSO₄, and the organics concentrated under reduced pressure. The crudeproduct was purified via medium pressure column chromatography using agradient solvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,followed by 3:97 methanol/CH₂Cl₂ as the solvent system to give 103 mg.Rf=0.22, 3:97 methanol/CH₂Cl₂, HPLC: Rt=15.29 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 147

Compound 147. To a solution of the resultant compound of Example 146B(80 mg, 0.16 mmol) in CH₂Cl₂ (5 mL) was added triethylamine (0.07 mL,0.48 mmol), followed by slow addition over 3 hours of the resultantcompound of Example 82A (53 mg, 0.19 mmol) as a solution in CH₂Cl₂ (3mL). The mixture was stirred at ambient temperature for 24 hours. Thesolution was diluted with 100 mL CH₂Cl₂ washed with water, dried overanhydrous MgSO₄, and the organics concentrated under reduced pressure.The crude product was purified via medium pressure column chromatographyusing a gradient solvent system of CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂, followed by 2:98 methanol/CH₂Cl₂ as the solvent systemto give 71.7 mg of the title compound. Rf=0.06, 3:97 methanol/CH₂Cl₂,HPLC: Rt=12.61 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 148

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=phenyl). A solution of 297 mg of the resultant compound of Example114B in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 217 mg ofbenzenesulfonyl chloride and 103 mg of sodium bicarbonate. The mixturewas stirred for 6 h, diluted with CH₂Cl₂, washed with saturated NaClthen dried over MgSO₄, filtered, and concentrated in vacuo to yield 426mg of the title product as a white solid. TLC: Rf=0.32, 5% diethylether/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=cyclopentylmethyl, E=phenyl, hydrochloridesalt). A solution of 400 mg of the resultant compound of Example 148A inethyl acetate was treated at −20° C. with HCl gas for 20 min, duringwhich time the temperature was allowed to warm to 20° C. Nitrogen wasthen bubbled through the mixture for 15 min and solvent removed in vacuoto yield 349 mg of white solid which was used directly for the ensuingreaction.

C. Compound 148. A solution of 40 mg of the resultant compound ofExample 148B in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, to a solution of 31 mg of the resultant compoundof Example 48A and 35 mg N,N-diisopropylethylamine in CH₂Cl₂. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaHCO₃ and saturated NaCl, then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 20% diethyl ether/CH₂Cl₂ as eluent to provide45 mg of the title product as a white solid. TLC: Rf=0.46, 20% diethylether/CH₂Cl₂. HPLC: Rt=15.78 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 149

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentymethyl,E=3-pyridyl). To a solution of 153 mg (0.422 mmol) of the resultantcompound of Example 114B in CH₂Cl₂ (4 mL) was added aqueous sodiumbicarbonate (1 mL), solid sodium bicarbonate 141.7 mg (1.69 mmol), andthe resultant compound of Example 144A 156.1 mg. (0.879 mmol). After 14h, the resulting mixture was diluted with CH₂Cl₂, washed with saturatedbrine, dried over magnesium sulfate, filtered and concentrated in vacuo.The residue was purified by flash chromatography using 20% to 40%EtOAc/CH₂Cl₂ eluent to provide 64.7 mg of the title product. TLC;Rf=0.24, 20% EtOAc/CH₂Cl₂.

B. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=3-pyridyl, hydrochloride salt). A solution of 273.1 mg (0.572 mmol) ofthe resultant compound of Example 149A in EtOAc (is mL) at −20° C. wastreated with anhydrous HCl gas for 10 min. The ice bath was removed andafter an additional 15 min., the reaction mixture was sparged withnitrogen then concentrated in vacuo. To a solution of the resultingresidue in CH₂Cl₂ (3 mL) was added, sequentially at ambient temperatureunder an atmosphere of nitrogen, with 0.076 mL (0.437 mmol)diisopropylethylamine and 34.3 mg (0.150 mmol) of the resultant compoundof Example 48A. The mixture was stirred for 16 h and then concentratedin vacuo. The residue was taken up in CH₂Cl₂ and washed with saturatedbrine, dried over magnesium sulfate, filtered, and concentrated invacuo. The residue was purified by low pressure silica gel columnchromatography using a gradient 20% to 50% EtOAc in CH₂Cl₂ eluent toyield 11.3 mg of the title product. TLC; Rf=0.15 40% EtOAc/CH₂Cl₂. HPLC:Rt=13.7 min; (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 150

A. 1-Piperidinesulfonyl chloride. A solution of 4 g of sulfuryl chloridein acetronitrile was treated dropwise with 861 mg of piperidine atambient temperature under an atmosphere of nitrogen. After completeaddition, the mixture was refluxed for 18 h, cooled to room temperatureand concentrated in vacuo to yield the title product as a red oil. TLC:Rf=0.86, CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=tert-butoxycarbonyl, D′isobutyl, E=piperidinyl). Asolution of 73 mg of the resultant compound of Example 39A in CH₂Cl₂ wastreated sequentially, at ambient temperature under an atmosphere ofnitrogen, with 121 mg of the resultant compound of Example 150A and 84mg of N,N-diisopropylethylamine. The mixture was stirred for 14 h,diluted with CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 5% diethyl ether/CH₂Cl₂ aseluent to provide 70 mg of the title product as a white solid. TLC:Rf=0.21 (5% diethyl ether in CH₂Cl₂). HPLC: Rt=17.40 min. (¹H)-NMR(CDCl₃) consistent with structure.

C. Compound XXII (A=H, D′=isobutyl, E=piperidinyl, hydrochloride salt).A solution of 70 mg of the resultant compound of Example 150B in ethylacetate was treated at −20° C. with HCl gas for 20 min during which timethe temperature was allowed to warm to 20° C. Nitrogen was then bubbledthrough the mixture for 15 min and solvent removed in vacuo to yield aviscous oil which was used directly for the ensuing reaction.

D. Compound 150. A solution of the resultant compound of Example 150C inCH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, to a solution of 50 mg of the resultant compound of Example48A and 56 mg N,N-diisopropylethylamine in CH₂Cl₂. the mixture wasstirred for 14 h, diluted with CH₂Cl₂, washed with saturated NaHCO₃ andsaturated NaCl, then dried over MgSO₄, filtered, and concentrated invacuo. The residue was purified by low pressure silica gelchromatography using 20% diethyl ether/CH₂Cl₂ as eluent to provide 16 mgof the title product as a white solid. TLC: Rf=0.45, (0 diethylether/CH₂Cl₂. HPLC: Rt=15.00 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 151

A. Compound XXII (A=tert butoxycarbonyl, D′=cyclopentylmethyl,E=4-trifluormethoxyphenyl). A solution of 71 mg of the resultantcompound of Example 114B in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ wastreated sequentially, at ambient temperature under an atmosphere ofnitrogen, with 76 mg of 4-trifluoromethoxybenzensulfonyl chloride and 25mg of sodium bicarbonate. The mixture was stirred, 14 h, diluted withCH₂Cl₂, washed with saturated NaCl then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 5% diethyl ether/CH₂Cl₂ as eluent to provide 92mg of the title product as a white solid. TLC: Rf=0.34, 5% diethylether/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=cyclopentylmethyl, E=4-trifluormethoxyphenyl,hydrochloride salt). A solution of 92 mg of the resultant compound ofExample 151A in ethyl acetate was treated at −20° C. with HCl gas for 20min, during which time the temperature was allowed to warm to 20° C.Nitrogen was then bubbled through the mixture for 15 min and solventremoved in vacuo to yield 83 mg of white solid which was used directlyfor the ensuing reaction.

C. Compound 151. A solution of 22 mg of the resultant compound ofExample 151B in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, to a solution of 15 mg of the resultant compoundof Example 48A and 16 mg N,N-diisopropylethylamine in CH₂Cl₂. Themixture was stirred for 60 h, diluted with CH₂Cl₂, washed with saturatedNaHCO₃ and saturated NaCl, then dried over MgSO₄ filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 20% diethyl ether/CH₂Cl₂ as eluent to provide23 mg of the title product as a white solid. TLC: Rf=0.44, 20% diethylether/CH₂Cl₂. HPLC: Rt=16.99 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 152

A. Compound XXII (A=tert-butoxycarbonyl, D′=isobutyl,E=4-trifluormethoxyphenyl). A solution of 97 mg of the resultantcompound of Example 39A in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ wastreated sequentially, at ambient temperature under an atmosphere ofnitrogen, with 113 mg of 4-trifluoromethoxybenzenesulfonyl chloride and36 mg of sodium bicarbonate. The mixture was stirred for 14 h, dilutedwith CH₂Cl₂, washed with saturated NaCl then dried over MgSO₄, filtered,and concentrated in vacuo. The residue was purified by low pressuresilica gel chromatography using 5% diethyl ether/CH₂Cl₂ as eluent toprovide 120 mg of the title product as a white solid. TLC: Rf=0.34, 5%diethyl ether/CH₂Cl₂. HPLC: Rt=18.54 min. (¹H)-NMR (CDCl₃) consistentwith structure.

B. Compound XXII (A=H, D′=isobutyl, E=4-trifluormethoxyphenyl,hydrochloride salt). A solution of 100 mg of the resultant compound ofExample 152A in ethyl acetate was treated at −20° C. with HCl gas for 20min, during which time the temperature was allowed to warm to 20° C.Nitrogen was then bubbled through the mixture for 15 min and solventremoved in vacuo to yield 89 mg of white solid which was used directlyfor ensuing reaction.

C. Compound 152. A solution of 41 mg of the resultant compound ofExample 152B in CH₂Cl₂ was added, at ambient temperature under theatmosphere of nitrogen, to a solution of 28 mg of the resultant compoundof Example 48A and 32 mg N,N-diisopropylethylamine in CH₂Cl₂. Themixture was stirred 14 h, diluted with CH₂Cl₂, washed with saturatedNaHCO₃ and saturated NaCl, then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 5% diethyl ether/CH₂Cl₂ as eluent to provide 30mg of the title product as white solid. TLC: Rf−0.08 (5% diethylether/CH₂Cl₂). HPLC: Rt=16.52 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 153

A. Compound XXII (A=tert-butoxycarbonyl, D′isobutyl, E=4-methoxyphenyl).To a solution of the resultant compound of Example 39A (600 mg, 1.77mmol) in CH₂Cl₂ (10 mL) was added 4-methoxybenzenesulfonyl chloride(0.55 g. 2.66 mmol) followed by the addition of a saturated solution ofsodium bicarbonate (3 mL) and 0.30 g of solid sodium bicarbonate. Themixture was stirred at ambient temperature overnight. The solution wasdiluted with 200 mL CH₂Cl₂, the organics were separated, dried overanhydrous MgSO₄, and the organics concentrated under reduced pressure.The crude product was purified via medium pressure liquid chromatographyusing a gradient solvent system of CH₂Cl₂ followed by 5:95 ether/CH₂Cl₂solution to give 630 mg of the title compound as a white solid. TLC:Rf=0.48, 3:97 methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent withstructure.

B. Compound XXII (A=H, D′=isobutyl, E=4-methoxyphenyl, hydrochloridesalt). To a solution of the resultant compound of Example 153A (0.63 g,1.24 mmol) in EtAc (5 mL) was added 30% w/w HCl in EtOAc (5 mL.) Themixture was stirred for 6 hours ambient temperature. The solution wasconcentrated under reduced pressure to give 0.59 g of a white solidwhich was used directly for subsequent reaction. TLC Rf=0.12, 3:97methanol/CH₂Cl₂.

C. Compound XXII (A=(3-pyridyl)-methyloxycarbonyl, D′=isobutyl,E=4-methoxyphenyl). To a solution of the resultant compound of Example153B (100 mg, 0.23 mmol) in CH₂Cl₂ (5 mL) was added triethylamine (0.1mL, 0.72 mmol) followed by slow addition over 3 hours of the resultantcompound of Example 82A (75 mg, 0.27 mmol) as a solution in CH₂Cl₂ (5mL). The mixture was stirred at ambient temperature for 24 hours. Theorganics was concentrated under reduced pressure and the crude productwas purified via medium pressure column chromatography using a gradientsolvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂, followed by3:97 methanol/CH₂Cl₂ as the solvent system to give 49.3 mg of the titlecompound. Rf=0.33, 3:97 methanol/CH₂Cl₂. HPLC: Rt=13.18 min, (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 154

Compound 154. To a solution of the resultant compound of Example 153B(100 mg, 0.20 mmol) in CH₂Cl₂ (5 mL) was added triethylamine (0.25 mL,1.8 mmol) followed by allylchloroformate (0.1 mL, 0.94 mmol). Themixture was stirred at ambient temperature for 24 hours. The solutionwas concentrated under reduced pressure and the crude product purifiedvia medium pressure column chromatography using a gradient solventsystem of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂ as the solvent systemto give 94 mg of the title compound. Rf=0.71, 3:97 methanol/CH₂Cl₂.HPLC: Rt=16.12 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 155

A. N-hydroxysuccinimidyl-1-methoxypropane-3-carbonate. A solution of 355mg of 2-methylene-1,3-propanediol in acetonitrile (30 mL) was addedsequentially, at ambient temperature, 65 mg of sodium hydride and 0.25mL iodomethane. The mixture was stirred for 12 h and concentrated invacuo. The residue was then taken up in 15 mL of acetonitrile andtreated sequentially, at ambient temperature under an atmosphere ofnitrogen, with 1.3 g of N,N-disuccinimidyl carbonate and 1.6 mL oftriethylamine. After stirring for 14 h, the reaction mixture wasconcentrated in vacuo and the residue was diluted CH₂C₂, washed withsaturated sodium bicarbonate solution and saturated brine, dried overmagnesium sulfate, filtered and concentrated in vacuo. The residue waspurified by silica gel chromatography with EtOAc as eluant to give 95 mgof the title compound. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 155. A solution of 0.056 mmol of the resultant compound ofExample 40A was reacted with the resultant compound of Example 155A inthe manner described in Example 132. After concentration of the mixturein vacuo and workup, the residue was purified by thick layer silica gelchromatography using 7% MeOH/CH₂Cl₂ as eluant followed by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100% CH₃CN/H₂Owith 0.1% TFA as eluant to obtain 3.7 mg of the title compound. TLC:Rf=0.45, 7% MeOH/CH₂Cl₂. HPLC: Rt=13.78 min.

EXAMPLE 156

A. 1-acetylindoline-5-sulfonyl chloride. A 1.02 g portion of1-acetylindoline was treated with 2 mL of chlorosulfonic acid at 0° C.The mixture was heated at 60° C. for 2 h, then treated with crushed ice,filtered and dried to give 1.3 g of the title compound which was useddirectly for subsequent reaction. TLC: Rf=0.18, 50% EtOAc/hexane.(¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=5-(N-acetyl)-indoline). To a solution of 60 mg of the resultantcompound of Example 114B in 15 mL of CH₂Cl₂ was added (5 mL) saturatedaqueous sodium bicarbonate solution, 50.0 mg sodium bicarbonate, and 60mg of the resultant compound of Example 156A. After 4 h, the resultingmixture was diluted with CH₂Cl₂, washed with saturated brine, dried overmagnesium sulfate and filtered. The mixture was then concentrated invacuo to give the desired product which was used directly for subsequentreaction. (¹H)-NMR (CDCl₃) consistent with structure.

C. Compound 156. A solution of 37 mg of the resultant compound ofExample 156B in EtOAc (15 mL) at 0° C. was treated with anhydroushydrogen chloride gas for 10 min., and allowed to stand for 12 h whilewarming to ambient temperature. This crude material was then reactedwith allyl chloroformate in the manner described in Example 87B. Afterconcentration of the mixture in vacuo and workup, the residue waspurified by thick layer silica gel chromatography using 7% MeOH/CH₂Cl₂as eluant followed by preparative reversed-phase C₁₈ HPLC using a lineargradient of 35% to 100% CH₃CN/H₂O with 0.1 TFA as eluant to obtain 10.5mg of the title compound. TLC: Rf=0.75, 10% MeOH/CH₂Cl₂. HPLC: Rt=15.78min; (¹H)-NMR (CDCL₃) consistent with structure.

EXAMPLE 157

Compound 157. A solution of 37 mg of the resultant compound of Example156B in EtOAc (15 mL) at 0° C. was treated with anhydrous hydrogenchloride gas for 10 min., and allowed to stand for 12 h while warming toambient temperature. This crude material was then reacted with theresultant compound of Example 48A in the manner described in Example 88.After concentration of the mixture in vacuo, the residue was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant to obtain 17.9 mg of the titlecompound. TLC: Rf=0.6, 10% MeOH/CH₂Cl₂. HPLC: Rt=14.68 min; (¹H)-NMR(CDCL₃) consistent with structure.

EXAMPLE 158

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl, E=H). To asolution of compound XX (A=Boc) (5.0 mmol) in ethanol (20 mL) was addedcyclohexylmethylamine (3.25 mL, 2.83 mmol) and the mixture was stirredfor 3 hours at ambient temperature. The solution was filtered andconcentrated under reduced pressure to give 1.49 g of a white solidwhich was used directly for subsequent reaction. TLC: Rf=0.14, 3:97methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,E=4-methoxyphenyl). To a solution of the resultant compound of Example158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added4-methoxybenzenesulfonyl chloride (0.66 g, 3.1 mmol) followed byaddition of a saturated solution of sodium bicarbonate (3 mL) and 0.18 gof solid sodium bicarbonate. The mixture was stirred at ambienttemperature overnight. The solution was diluted with 200 mL CH₂Cl₂, theorganics separated, dried over anhydrous MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂ as the solvent system to give 340 mg of the titlecompound as a white solid. TLC: Rf=0.39, 3:97 methanol/CH₂Cl₂, (¹H)-NMR(CDCL₃) consistent with structure.

C. Compound XXI (A=H, D′=cyclohexylmethyl, E=4-methoxyphenyl,hydrochloride salt). To a solution of the resultant compound of Example158B (0.34 g, 0.62 mmol) in EtOAc (10 mL) was added 30% w/w HCl in EtOAc(5 mL). The mixture was stirred for 3 hours at ambient temperature. Thesolution was concentrated under reduced pressure to give 0.3 g of awhite solid which was used directly for subsequent reaction. TLC:Rf=0.12, 3:97 methanol/CH₂Cl₂.

D. Compound 158. To a solution of the resultant compound of Example 158C(100 mg, 0.21 mmol) in CH₂Cl₂ (8 mL) was added triethylamine (0.2 mL,1.44 mmol) followed by the resultant compound of Example 48A (71 mg,0.31 mmol). The mixture was stirred at ambient temperature for 6 hours.The solution was diluted with CH₂Cl₂, (200 mL) washed with a saturatedsolution of sodium bicarbonate (30 mL), the organics separated, driedover anhydrous MgSO₄ and concentrated under reduced pressure and thecrude product purified via medium pressure column chromatography using agradient solvent system of CH₂Cl₂ followed by 10:90 EtOAc/CH₂Cl₂ as thesolvent system to give 84.9 mg of the title compound. TLC: Rf=0.48, 3:97methanol/CH₂Cl₂, HPLC: Rt=16.35 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 159

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,E=4-fluorophenyl). To a solution of the resultant compound of Example158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added4-fluorobenzenesulfonyl chloride (0.62 g, 3.2 mmol) followed by additionof a saturated solution of sodium bicarbonate (3 mL) and 0.18 g of solidsodium bicarbonate. The mixture was stirred at ambient temperatureovernight. The solution was diluted with 200 mL CH₂Cl₂, the organicsseparated, dried over anhydrous MgSO₄, and the organics concentratedunder reduced pressure. The crude product was purified via mediumpressure liquid chromatography using CH₂Cl₂ followed by 1:99methanol/CH₂Cl₂ solution as the solvent system to give 280 mg of a whitesolid. TLC: Rf=0.47, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistentwith structure.

B. Compound XXII (A=H, D′=cyclohexylmethyl, E=4-fluorophenyl,hydrochloride salt). To a solution of the resultant compound of Example159A (0.28 g, 0.52 mmol) was added 30% w/w HCl in EtOAc (10 mL). Themixture was stirred for 3 hours at ambient temperature. The solution wasconcentrated under reduced pressure to give 0.23 g of a white solidwhich was used directly for subsequent reaction. TLC: Rf=0.13, (3:97methanol/CH₂Cl₂, (¹H)-NMR (CDCL₃) consistent with structure.

C. Compound 159. To a solution of the resultant compound of Example 159C(100 mg, 0.21 mmol) in CH₂Cl₂ (8 mL) was added triethylamine (0.2 mL,1.44 mmol) followed by the resultant compound of Example 48A (73 mg,0.32 mmol). The mixture was stirred at ambient temperature for 6 hours.The solution was diluted with CH₂Cl₂, (200 mL) washed with saturatedsolution of sodium bicarbonate (30 mL), dried over anhydrous MgSO₄, theorganics concentrated under reduced pressure and the crude productpurified via medium pressure column chromatography using a gradientsolvent system of CH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂ as the solventsystem to give 54 mg of the title compound. TLC: Rf=0.46, 3:97methanol/CH₂Cl₂, HPLC: Rt=16.48 min; (1H)-NMR (CDCL₃) consistent withstructure.

EXAMPLE 160

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclohexylmethyl,E=4-acetamidophenyl). To a solution of the resultant compound of Example158A (400 mg, 1.06 mmol) in CH₂Cl₂ (10 mL) was added4-acetamidobenzenesulfonyl chloride (0.75 g, 3.2 mmol) followed byaddition of a saturated solution of sodium bicarbonate (3 mL) and 0.18 gof solid sodium bicarbonate. The mixture was stirred at ambienttemperature overnight. The solution was diluted with 200 mL CH₂Cl₂, theorganics separated, dried over anhydrous MgSO₄, and the organicsconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂ and 2:98 methanol/CH₂Cl₂ as the solvent system to give290 mg of the title compound as a white solid. TLC: Rf=0.14, 3:97methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=cyclohexylmethyl, E=4-acetamidophenyl,hydrochloride salt). To the resultant compound of Example 160A (0.29 g,0.51 mmol) was added 30% w/w HCl in EtOAc (10 mL). The mixture wasstirred for 3 hours at ambient temperature. The solution wasconcentrated under reduced pressure to give 0.28 g of a white solidwhich was used directly for subsequent reaction. TLC: Rf=0.10, 3:97methanol/CH₂Cl₂.

C. Compound 160. To a solution of the resultant compound of Example 160B(100 mg, 0.20 mmol) in CH₂Cl₂ (8 mL) was added triethylamine (0.2 mL,1.44 mmol) followed by the resultant compound of Example 48A (67 mg,0.30 mmol). The mixture was stirred at ambient temperature for 6 hours.The solution was diluted with CH₂Cl₂, (200 mL) washed with saturatedsolution of sodium bicarbonate (30 mL), dried over anhydrous MgSO₄, theorganics concentrated under reduced pressure and the crude productpurified via medium pressure column chromatography using a gradientsolvent system of CH₂Cl₂, followed by 10:90 EtOAc/CH₂Cl₂, followed by20:80 EtOAc/CH₂Cl₂ as the solvent system to give 56.8 mg of a whitesolid. TLC: Rf=0.17, 3:97 methanol/CH₂Cl₂, HPLC: Rt=14.65 min; (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 161

A. 4-Morpholinesulfonyl chloride. A solution of 4.6 g of sulfurylchloride in acetonitrile was treated dropwise with 996 mg of morpholineat ambient temperature under an atmosphere of nitrogen. After completeaddition, the mixture was refluxed for 16 h, cooled to room temperature,and concentrated in vacuo to yield the title product as a red oil. TLC:Rf=0.65 CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A-tert-butoxycarbonyl, D′=isobutyl, E=morpholinyl). Asolution of 98 mg of the resultant compound of Example 39A in 4:1CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, at ambienttemperature under an atmosphere of nitrogen, with 270 mg of theresultant compound of Example 161A and 122 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, dried over MgSO₄,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using CH₂Cl₂ as eluent followed bypreparative HPLC to provide 22 mg of the title product as an oily solid.TLC: Rf=0.46, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.50 min. (¹H)-NMR(CDCl₃) consistent with structure.

C. Compound XXII (A=H, D′=isobutyl, E=morpholinyl, hydrochloride salt).A solution of 22 mg of the resultant compound of Example 161B in ethylacetate was treated at −20° C. Nitrogen was then bubbled through themixture for 15 min and solvent removed in vacuo to yield an oilysemi-solid mass which was used directly for the ensuing reaction.

D. Compound 161. A solution of the resultant compound of Example 161C inCH₂Cl₂ was added, at ambient temperature under an atmosphere ofnitrogen, to a solution of 16 mg of the resultant compound of Example48A and 18 mg N,N-diisopropylethylamine in CH₂Cl₂. The mixture wasstirred for 14 h, diluted with CH₂Cl₂, washed and saturated with NaHCO₃and saturated NaCl, then dried over MgSO₄, filtered, and concentrated invacuo. The residue was purified by preparative HPLC to provide 21 mg ofthe title product as an oily solid. TLC: Rf=0.22, 20% diethylether/CH₂Cl₂. HPLC: Rt=13.01 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 162

Compound 162. A solution of 30 mg of the resultant compound of Example166A was deprotected with hydrogen chloride gas and the resultantcompound was reacted with the resultant compound of Example 155A in themanner described in Example 155B. After concentration of the mixture invacuo and workup, the residue was purified by thick layer silica gelchromatography using 5% MeOH/CH₂Cl₂ as eluant, followed by preparativereversed-phase C₁₈ HPLC using a linear gradient of 35% to 100%CH₃CN/H₂O) with 0.1% TFA as eluant to obtain 6.2 mg of the titlecompound. TLC: Rf=0.65, 5% MeOH/CH₂Cl₂. HPLC: Rt=15.93 min (H-NMR(CDCl₃) consistent with structure.

EXAMPLE 163

Compound 163. A 120.3 mg portion of the resultant compound of Example153B was reacted with the resultant compound of Example 82A as describedin Example 82B. After workup and concentration in vacuo, the residue waspurified by low pressure silica gel column chromatography using 50%EtOAc in CH₂Cl₂ eluent, followed by preparative reversed-phase C₁₈ HPLCusing a linear gradient of 40% to 100% acetonitrile/water for elution toobtain 44.3 mg of the title compound. TLC: Rf=0.18, 50% EtOAc/CH₂Cl₂.HPLC: Rt=13.13 min; (¹H) NMR (CDCl₃) consistent with structure.

EXAMPLE 164

A. N-hydroxysuccinimidyl-(2-phenyl)ethyl carbonate.

A solution of 306 mg of phenethyl alcohol and 535 mg ofN,N′-disuccinimidyl carbonate in acetonitrile was treated, at ambienttemperature under an atmosphere of nitrogen, with 810 mg ofN,N-diisopropylethylamine. The mixture was stirred for 60 h andconcentrated in vacuo. The residue was taken up in ethyl acetate andwashed with saturated NaHCO₃, saturated NaCl, then dried over MgSO₄,filtered, and concentrated in vacuo to yield the title product as ayellow oil. TLC: Rf=0.40 (5% methanol in CH₂Cl₂). (¹H)-NMR (CDCl₃)consistent with structure.

B. Compound 164. A solution of 81 mg of the resultant compound ofExample 164A in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, to a solution of 41 mg of the resultant compoundof example 40a and 45 mg N,N-diisopropylethylamine in CH₂Cl₂. Themixture was stirred for 4 h, diluted with CH₂Cl₂, washed with saturatedNaHCO₃ and saturated NaCl, then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was subjected to preparative HPLC toyield 18 mg of the title product. TLC: Rf=0.83 (5:10:85NH₄H/CH₃OH/CH₂Cl₂). HPLC: Rt=15.78 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 165

Compound 165. A solution of 36 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 20 mg ofp-toluenesulfonyl chloride and 18 mg of sodium bicarbonate. The mixturewas stirred for 3 h, diluted with CH₂Cl₂, washed with saturated NaClthen dried over MgSO₄, filtered, and concentrated in vacuo. The residuewas purified by low pressure silica gel chromatography using 5% diethylether/CH₂Cl₂ as eluent to provide 38 mg of the title product as a whitesolid. TLC: Rf=0.15, 5% diethyl ether/CH₂Cl₂. HPLC: Rt=15.27 min.(¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 166

A. Compound XXII (P=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=4-methoxyphenyl). To a solution of the resultant compound of Example114B (1.8 g, 4.96 mmol) in CH₂Cl₂ (10 mL) was added4-methoxylbenzensulfonyl chloride (2.10 g, 9.93 mmol), followed byaddition of a saturated solution of sodium bicarbonate (3 mL) and 0.83 gof solid sodium bicarbonate. The mixture was stirred at ambienttemperature for 24 hours. The solution was diluted with 200 mL CH₂Cl₂,the organics were separated, dried over anhydrous MgSO₄, andconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂ followed by 2:98 methanol/CH₂Cl₂ as the solvent systemto give 1.49 g of the title compound as a white solid. TLC: Rf=0.37,3:97 methanol/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (P═H, D′=cyclopentylmethyl, E=4-hydroxyphenyl). Asolution of the resultant compound of Example 166A (1.11 g, 2.08 mmol)in CH₂Cl₂ (20 mL) was added to a solution of boron tribromide in CH₂Cl₂(1.0 M, 10.4 mL). The mixture was stirred at ambient temperature for 24hours. The solution was poured onto 40 mL of a saturated solution ofsodium bicarbonate. The aqueous layer was extracted with 250 mL CH₂Cl₂followed by extraction with 250 mL EtOAc. The combined organics weredried over anhydrous MgSO₄, concentrated under reduced pressure and thecrude product purified via medium pressure column chromatography using agradient solvent system of CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,followed by 9:98 methanol/CH₂Cl₂, followed by a 1:5:95 concentratedNH₄OH/methanol/CH₂Cl₂ solution as the solvent system to give 0.38 g ofthe title compound. TLC: Rf=0.18, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃)consistent with structure.

C. Compound 166. To a solution of the resultant compound of Example 166B(300 mg, 0.69 mmol) in CH₂Cl₂ (5 mL) was added triethylamine (0.12 mL,8.6 mmol), followed by slow addition over 3 hours of the resultantcompound of Example 82A (0.21 g, 0.77 mmol) as a solution in CH₂Cl₂ (5mL). The mixture was stirred at ambient temperature for 24 hours. Thesolution was diluted with 250 mL CH₂Cl₂, washed with water, dried overanhydrous MgSO₄, and the organics concentrated under reduced pressure.The crude product was purified via medium pressure column chromatographyusing a gradient solvent system of CH₂Cl₂ followed by 1:99methanol/CH₂Cl₂, followed by 2:98 methanol/CH₂Cl₂ as the solvent systemto give 110 mg of a white solid. TLC: Rf=0.14 (3:97 methanol/CH₂Cl₂),HPLC: Rt=12.69 min, (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 167

Compound 167. A solution of 102 mg of the resultant compound of Example51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 65 mg ofp-nitrobenzenesulfonyl chloride and 51 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaCl, then dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by low pressure silica gel chromatography using 20%diethyl ether/CH₂Cl₂ as eluent to provide 124 mg of the title product asa white solid. TLC: Rf=0.36, 20% diethyl ether/CH₂Cl₂. HPLC: Rt=15.15min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 168

Compound 168. A solution of 124 mg of the resultant compound of Example167 in ethyl acetate was treated, at ambient temperature, with 13 mg of10% palladium on carbon. The mixture was stirred for 14 h under anatmosphere of hydrogen, filtered through a pad of Celite filter agent,and concentrated in vacuo. The residue was subjected to preparative HPLCto yield 82 mg of the title product as a white solid. TLC: Rf=0.10, 20%ether/CH₂Cl₂. HPLC: Rt=13.16 min. (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 169

Compound 169. To a solution of the resultant compound of Example 166B(80 mg, 0.18 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution ofsodium bicarbonate (5 mL) followed by the addition of the resultantcompound of Example 48A (55 mg, 0.24 mmol). The mixture was stirred atambient temperature for 5 hours. The solution was diluted with 200 mLCH₂Cl₂, the organics separated, dried over anhydrous MgSO₄, andconcentrated under reduced pressure. The crude product was purified viamedium pressure liquid chromatography using CH₂Cl₂, followed by 1:99methanol/CH₂Cl₂ as the solvent system to give 56 mg of the titlecompound as a white solid. TLC: Rf=0.24, 3:97 methanol/CH₂Cl₂, HPLC:Rt=14.29 min. (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 170

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl, E=4,nitrophenyl). To a solution of the resultant compound of Example 114B(250 mg, 0.69 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution ofsodium bicarbonate (5 mL) followed by solid sodium bicarbonate (0.12 g,1.37 mmol) and 4-nitrobenzensulfonyl chloride (200 mg, 0.9 mmol). Themixture was stirred at ambient temperature for 24 hours. The solutionwas diluted with 200 mL CH₂Cl₂, the organics separated, dried overanhydrous MgSO₄, and concentrated under reduced pressure. The crudeproduct was purified via medium pressure liquid chromatography using agradient solvent system of CH₂Cl₂ followed by 1:99 methanol/CH₂Cl₂ togive 360 mg of the title compound as an orange solid. TLC: Rf=0.45, 3:97methanol/CH₂Cl₂. (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound XXII (A=H, D′=cyclopentylmethyl, E-4-nitrophenyl,hydrochloride salt). To the resultant compound of Example 170A (360 mg,0.66 mmol) was added 10% w/w HCl in EtOAc (15 mL). The mixture wasstirred for 3 hours at ambient temperature. The solution wasconcentrated under reduced pressure to give 310 mg of the title compoundas an orange solid which was used directly for subsequent reaction. TLC:Rf=0.70, 1:10:90 NH₄OH/methanol/CH₂Cl₂.

C. Compound 170. To a solution of the resultant compound of Example 170B(310 mg, 0.64 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution ofsodium bicarbonate (5 mL) followed by the addition of solid sodiumbicarbonate (0.11 g, 1.3 mmol) and the resultant compound of Example 48A(0.18 g, 0.77 mmol). The mixture was stirred at ambient temperature for24 hours. The solution was diluted with 150 mL CH₂Cl₂, the organicsseparated, dried over anhydrous MgSO₄, and concentrated under reducedpressure. The crude product was purified via medium pressure liquidchromatography using CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂ as thesolvent system to give 0.32 g of the title compound as a white solid.TLC: Rf=0.28, 3:97 methanol/CH₂Cl₂, HPLC: Rt=16.06 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 171

Compound 171. A solution of the resultant compound of Example 170C (0.19g, 0.34 mmol) in EtOAc (10 mL) was treated at ambient temperature with50 mg of 10% palladium on carbon and hydrogenated for 72 hours under aslight positive pressure of hydrogen. The mixture was filtered andconcentrated in vacuo and the crude product purified via medium pressureliquid chromatography using CH₂Cl₂, followed by 1:99 methanol/CH₂Cl₂,followed by 3:97 methanol/CH₂Cl₂, followed by 10:90 methanol/CH₂Cl₂ asthe solvent system to give 97 mg of the title compound as a white solid.TLC: Rf=0.25, 3:97 methanol/CH₂Cl₂, HPLC: Rt=14.28 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 172

A. Compound XXII (A=tert-butoxycarbonyl, D′=cyclopentylmethyl,E=2,4-dinitrophenyl). To a solution of the resultant compound of Example114B (500 mg, 1.38 mmol) in CH₂Cl₂ (15 mL) was added a saturatedsolution of sodium bicarbonate (5 mL) followed by solid sodiumbicarbonate (0.23 g, 2.76 mmol) and 2,4-dinitrobenzenesulfonyl chloride(440 mg, 1.65 mmol). The mixture was stirred at ambient temperature for2 hours. The solution was diluted with 200 mL CH₂Cl₂, the organicsseparated, dried over anhydrous MgSO₄, and concentrated under reducedpressure. The crude product was purified via medium pressure liquidchromatography using a gradient solvent system of CH₂Cl₂, followed by1:99 methanol/CH₂Cl₂ to give 700 mg of the title compound as a brownsolid. TLC: Rf=0.48, 3:97 methanol/CH₂Cl₂, (¹H)-NMR (CDCl₃), consistentwith structure.

B. Compound XXII (A=H, D′=cyclopentylmethyl, E-2,4-dinitrophenyl,hydrochloride salt). To a the resultant compound of Example 172A (700mg, 1.18 mmol) was added 10% w/w HCl in EtOAc (20 mL). The mixture wasstirred for 3 hours at ambient temperature. The solution wasconcentrated under reduced pressure to give 590 mg of the title compoundas a brown solid which was used without subsequent purification. TLC:Rf=0.55, 1:10:90 NH₄OH/methanol/CH₂Cl₂.

C. Compound 172. To a solution of the resultant compound of 172B (590mg, 1.11 mmol) in CH₂Cl₂ (15 mL) was added a saturated solution ofsodium bicarbonate (5 mL), followed by solid sodium bicarbonate (0.19 g,2.2 mmol) and the resultant compound of Example 48A (0.31 g, 1.3 mmol).The mixture was stirred at ambient temperature for 24 hours. Thesolution was diluted with 150 mL CH₂Cl₂, the organics separated, driedover anhydrous MgSO₄, and the organics concentrated under reducedpressure. The crude product was purified via medium pressure liquidchromatography using a CH₃OH/CH₂Cl₂ gradient as eluant, to yield theproduct as 0.59 g of a white solid. HPLC: Rt=16.36 min, (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 173

Compound 173. A solution of the resultant compound of Example 172C (0.20g, 0.33 mmol) in EtOAc (10 mL) was treated under ambient temperaturewith 50 mg of 10% palladium on carbon and hydrogenated for 72 hoursunder a slight positive pressure of hydrogen. The mixture was filteredand concentrated in vacuo and the crude product purified via mediumpressure liquid chromatography using CH₂C₂, followed by 1:99;methanol/CH₂Cl₂, 3:97 methanol/CH₂Cl₂, and 10:90 methanol/CH₂Cl₂ as thesolvent system to give 120.2 mg of the title compound as a light brownsolid. TLC: Rf=0.17, 3:97 methanol/CH₂Cl₂, HPLC: Rt=13.47 min, (¹H)-NMR(CDCl₃) consistent with structure.

EXAMPLE 174

A. 4-Benzyloxybenzenesulfonyl chloride. To 0.87 g of dimethylformamide,at 0° C. under an atmosphere of nitrogen, was added 1.61 g of sulfurylchloride. The mixture was stirred for 15 min and treated with 2.00 g ofbenzyl phenyl ether. The mixture was then heated at 100° C. for 1.5 h,cooled to about 40° C., poured onto ice, extracted with CH₂Cl₂, driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 10% ethylacetate in hexane as eluent to provide 0.78 g of the title product as awhite solid. TLC: Rf=0.46, 10% ethyl acetate in hexane. (¹H)-NMR (CDCl₃)consistent with structure.

B. Compound 174. A solution of 30 mg of the resultant compound ofExample 51D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treatedsequentially, at ambient temperature under an atmosphere of nitrogen,with 24 mg of the resultant compound of Example 174A and 18 mg of sodiumbicarbonate. The mixture was stirred for 14 h, diluted with CH₂Cl₂,washed with saturated NaCl then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 20% diethyl ether/CH₂Cl₂ as the eluent toprovide 14 mg of the title product as a white solid. TLC: Rf=0.43, 20%diethyl ether/CH₂Cl₂. HPLC: Rt=17.01 min. (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 175

Compound 175. A solution of 11 mg of the resultant compound Example 174Bin ethyl acetate was treated at ambient temperature, with 2 mg of 10%palladium on carbon. The mixture was stirred for 14 h under anatmosphere of hydrogen, filtered through a pad of Celite filter agent,filtered, and concentrated in vacuo. The residue was purified by lowpressure silica gel chromatography using 10% methanol in CH₂Cl₂ as theeluent to provide 9 mg of the title product as a white solid. TLC:Rf=0.38, 10% methanol in CH₂Cl₂. HPLC: Rt=13.37 min. (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 176

A. 1,3-Benzodioxole-5-sulfonyl chloride. To 3.50 g of dimethylformamide,at 0° C. under an atmosphere of nitrogen, was added 6.47 g of sulfurylchloride. The mixture was stirred 15 min and treated with 5.32 g of1,3-benzodioxole. The mixture was then heated at 120° C. for 45 min,cooled to about 40° C., poured onto ice, extracted with CH₂Cl₂, driedover MgSO₄, filtered, and concentrated in vacuo. The residue waspurified by low pressure silica gel chromatography using 40% CH₂Cl₂ inhexane as eluent to provide 2.70 g of the title product as a yellowsolid. TLC: Rf=0.37, 40% CH₂Cl₂ in hexane. (¹H)-NMR (CDCl₃) consistentwith structure.

B. Compound XXII (A=tert-butoxy, D′=isobutyl, E=3,4-benzodioxole). Asolution of 49 mg of the resultant compound of Example 39A in 4:1CH₂Cl₂/saturated aqueous NaHCO₃ was treated sequentially, at ambienttemperature under an is atmosphere of nitrogen, with 45 mg of theresultant compound of Example 176A and 28 mg of sodium bicarbonate. Themixture was stirred for 14 h, diluted with CH₂Cl₂, washed with saturatedNaCl then dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was purified by low pressure silica gel chromatography using 20%diethyl ether/CH₂Cl₂ as the eluent to provide 71 mg of the title productas a waxy solid. TLC: Rf=0.65, 20% diethyl ether/CH₂Cl₂. (¹H)-NMR(CDCl₃) consistent with structure.

C. Compound XXII (A=H, D′=isobutyl, E=3,4-benzodioxole, hydrochloridesalt). A solution of 71 mg of the resultant compound of Example 176B inethyl acetate was treated at −20° C. with HCl gas. The HCl was bubbledthrough the mixture for 20 min over which time the temperature wasallowed to warm to 20° C. Nitrogen was then bubbled through the mixturefor 15 min and solvent removed in vacuo to yield 66 mg of the titleproduct as a white solid which was used directly in subsequentreactions.

D. Compound 176. A solution of 18 mg of the resultant compound ofExample 176C in CH₂Cl₂ was added, at ambient temperature under anatmosphere of nitrogen, to a solution of 13 mg of the resultant compoundof Example 48A and 14 mg N,N-diisopropylethylamine in CH₂Cl₂. Themixture was stirred for 16 h, diluted with CH₂Cl₂, washed with saturatedNaHCO₃ and saturated NaCl, then dried over MgSO₄, filtered, andconcentrated in vacuo. The residue was purified by low pressure silicagel chromatography using 5% diethyl ether/CH₂Cl₂ as the eluent toprovide 9 mg of the title product as a white solid. TLC: Rf=0.14, 5%diethyl ether 1/CH₂Cl₂. HPLC: Rt=15.52 min; (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 177

A. (4-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. To a solution of1.50 g of p-nitrophenyl chloroformate in 30 mL of CH₂Cl₂ at 0° C. wasadded sequentially, 0.77 mL of 4-methoxybenzyl alcohol and 0.82 mL of4-methyl morpholine. After stirring for a half hour at ambienttemperature, the resulting mixture was diluted with CH₂Cl₂, washed withwater, brine, dried over magnesium sulfate, filtered and concentrated invacuo to yield a pale yellow solid which was triturated withCH₂Cl₂/hexane and filtered to yield 1.51 g of the title compound. TLC:Rf=0.40, 20% EtOAc/hexane.

B. Compound 177. To a solution of 96.7 mg of the resultant compound ofExample 141A in 2 mL of CH₂Cl₂ was added sequentially, 90 μL ofdiisopropylethylamine and 81.3 mg of the resultant compound of Example178A. After stirring for 24 hours, the mixture was diluted with CH₂C₂,washed with water and brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by preparative thinlayer chromatography using 5% methanol in CH₂Cl₂ eluent to yield 104.8mg of the title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.66min, (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 178

A. (3-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. Prepared by thesame route as described for Example 177A, except 3-methoxybenzyl alcoholwas utilized for reaction with p-nitrophenyl chloroformate to yield thetitle compound as a pale yellow solid. TLC: Rf=0.40, 20% EtOAc/hexane.

B. Compound 178. To a solution of 97.8 mg of the resultant compound ofExample 141A in 2 mL of CH₂Cl₂ was added sequentially, 91 μL ofdisopropylethylamine and 82.2 mg of the resultant compound of Example178A. After stirring for 24 hours, the mixture was diluted with CH₂Cl₂,washed with water and brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by preparative thinlayer chromatography using 5% methanol in CH₂Cl₂ eluent to yield 25.7 mgof the title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.75min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 179

A. (2-Methoxyphenyl)-methyl-4-nitrophenyl carbonate. Prepared by thesame route as described for Example 177A, except 2-methoxybenzyl alcoholwas utilized for reaction with p-nitrophenyl chloroformate to yield thetitle compound as a pale yellow solid. TLC: Rf=0.40, 20% EtOAc/hexane.

B. Compound 179. To a solution of 97.8 mg of the resultant compound ofExample 141A in 2 mL of CH₂Cl₂ was added sequentially, 99 μL ofdiisoprophylethylamine and 89.2 mg of the resultant compound of Example179A. After stirring for 24 hours the mixture was diluted with CH₂Cl₂,washed with water and brine, dried over magnesium sulfate, filtered andconcentrated in vacuo. The residue was purified by preparative thinlayer chromatography using 5% methanol in CH₂Cl₂ eluent to yield 107.0mg of the title compound. TLC: Rf=0.4, 20% EtOAc/hexane, HPLC: Rt=17.58min. (¹H)NMR (CDCl₃) consistent with structure.

EXAMPLE 180

A. 2,3-Dihydrobenzofuran-5-sulfonyl chloride. To 3.35 g ofdimethylformamide, at 0° C. under an atmosphere of nitrogen, added 6.18g of sulfuryl chloride. The mixture was stirred 15 min and treated with4.69 g of 2,3-dihydrobenzofuran. The mixture was then heated at 100° C.for 1.5 h, cooled to about 40° C., poured onto ice, extracted withCH₂Cl₂, dried over MgSO₄, filtered, and concentrated in vacuo. Theresidue was taken up in ethyl acetate, cooled to 5° C. for 16 h, and theresultant pink crystals collected by vacuum filtration to provide 6.12 gof the title product. TLC: Rf=0.41, 10% ethyl acetate in hexane.(¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 180. A solution of 32 mg of the resultant compound ofExample 140D in 4:1 CH₂Cl₂/saturated aqueous NaHCO₃ was treatedsequentially, at ambient temperature under an atmosphere of nitrogen,with 22 mg of the resultant compound of Example 180A and 18 mg of sodiumbicarbonate. The mixture was stirred 14 h, diluted with CH₂Cl₂, washedwith saturated NaCl then dried over MgSO₄, filtered, and concentrated invacuo. The residue was purified by low pressure silica gelchromatography using 20% diethyl ether/CH₂Cl₂ as eluent to provide 20 mgof the title product as a white solid. TLC: Rf=0.52, 20% diethylether/CH₂Cl₂. HPLC: Rt=15.49 min (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 181

Compound 181. A solution of the resultant compound of Example 140D (150mg, 0.4 mmol) in CH₂Cl₂ (10 mL) was added a saturated solution of sodiumbicarbonate (5 mL) followed by solid sodium bicarbonate (0.1 g, 1.2mmol) and 4-cyanobenzensulfonyl chloride (0.1 g, 0.48 mmol). The mixturewas stirred at ambient temperature for 4 hours. The solution was dilutedwith 200 mL CH₂Cl₂, the organics separated, dried over anhydrous MgSO₄,and the organics concentrated under reduced pressure. The crude productwas purified via medium pressure liquid chromatography using CH₂Cl₂,followed by 1:99 methanol/CH₂Cl₂ solution as the solvent system to give0.19 g (86% yield) of the title compound as a white solid. TLC: Rf=0.40,3:97 methanol/CH₂Cl₂, HPLC: Rt=15.02 min, (¹H)-NMR (CDCl₃) consistentwith structure.

EXAMPLE 182

Compound 182. This compound was prepared from the resultant compound ofExample 114D and the resultant compound of Example 48A in the samemanner described in Example 88. After workup and purification bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA as eluant, 32.8 mg of the title compoundwas obtained. TLC: Rf=0.25, 4% MeOH/CH₂Cl₂. HPLC: Rt=16.06 min; (¹H)NMR(CDCl₃) consistent with structure.

EXAMPLE 183

Compound 183. This compound was prepared from the resultant compound ofExample 84 by treatment with hydrogen chloride gas and subsequentreaction with the resultant compound of Example 48A in the mannerdescribed in Example 132. After workup and purification bycrystallization from EtOAc, 33.0 mg of the title compound was obtainedas a white solid. TLC: Rf=0.25, 4% MeOH/CH₂Cl₂. HPLC: Rt=17.71 min;(¹H)-NMR (CDCL₃) consistent with structure.

EXAMPLE 184

A. (N-tert-butoxycarbonyl)-(R)-3-pyrrolidinyl-N-hydroxysuccinimidylcarbonate. To a solution of 1.0 g of (R)-3-hydroxypyrrolidine intetrahydrofuran (50 mL) was added sequentially, at ambient temperature,3.75 g of di-tert-butyl dicarbonate and 1 mL of 2N sodium hydroxide. Themixture was stirred for 1 hour, filtered and concentrated in vacuo. Theresultant compound was reacted with N,N-disuccinimidyl carbonate in themanner described in Example 155A. Workup and purification by thick layersilica gel chromatography using an EtOAc eluent yielded the titlecompound as a white solid; (¹H)-NMR (CDCl₃) consistent with structure.

B. Compound 184. A solution of 350 mg of the resultant compound ofExample 166A was deprotected with hydrogen chloride gas and theresultant compound was reacted with the resultant compound of Example184A in the manner described in Example 88. After concentration of themixture in vacuo and workup, the residue was purified by thick layersilica gel chromatography using 7% MeOH/CH₂Cl₂ as eluant, to obtain 120mg of the title compound. TLC: Rf=0.45, 5% MeOH/CH₂Cl₂. HPLC: Rt=16.97min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 185

Compound 185. A solution of 120 mg of the resultant compound of Example184B in EtOAc (25 mL) at 0° C. was treated with anhydrous hydrogenchloride gas for 10 min., and allowed to stand for 12 h while warming toambient temperature. Concentration in vacuo yielded 110 mg of the titlecompound. TLC: Rt=0.35, 10% MeOH/89% CH₂Cl₂/1% NH₄OH. HPLC: Rt=13.72min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 186

A. Compound XXX ((syn, anti)-OH, A=carbobenzyloxy, R³=(s)-sec-butyl,R^(3′)═H, D′-benzyl, A′=tert-butoxycarbonyl). A solution of 1.37 g ofthe resultant compound of Example 1B in 150 mL of methylene chloride wastreated with 1.03 g of Cbz-Ile, 523 mg of HOBT.H₂O, and 742 mg of EDC.The mixture was stirred for 18 h, then diluted with 3 volumes of diethylether and washed sequentially with water, saturated NaHCO₃ solution, 10%KHSO₄ solution, and brine. After drying over MgSO₄ and concentrating invacuo, the residue was purified by chromatography on a silica gel columnusing a gradient of 1% to 1.5% MeOH in CH₂Cl₂ as eluant to yield 2.10 gof the title compound as a white foam. TLC: Rf=0.51, 5% methanol/CH₂Cl₂.

B. Compound XXX ((syn, anti)-OH, A=carbobenzyloxy, R³=(S)-sec-butyl,R³═H, D′=benzyl, A′=H), hydrochloride salt. A solution of 650 mg of theresultant compound of Example 12A in 12 mL of ethyl acetate was cooledin an ice/water bath and treated with a slow stream of HCl gas forapproximately 6 min with vigorous stirring. The mixture was capped andstirred for an additional 10 min, then purged with a stream of nitrogenfor 15 minutes and concentrated in vacuo to yield a white solid whichwas used without subsequent purification. TLC: Rf=0.18, 95:5:0:5CH₂Cl₂/methanol/concentrated NH₄OH.

C. Compound 186. A solution of 20 mg of the resultant compound ofExample 186B in 0.8 mL of methylene chloride was cooled in ice/methanol(approximately 15° C.), then treated with 13.8 μL of DIEA followed by7.6 mg of α-toluene sulfonyl chloride. The mixture was stirred for 15 h,warming slowly to ambient temperature. The mixture was concentrated to asmall volume, applied to a 0.5 mm thick prep plate and eluted with 3.5%MeOH/CH₂Cl₂. The band containing the desired diastereomer was isolatedand eluted with 8% MeOH/CH₂Cl₂ to yield 4.8 mg of the title compound.TLC: Rf=0.42, 15% diethyl ether/CH₂Cl₂. HPLC: Rt=17.81 min. NMR (CDCl₃):0.78 (dd, 6H) 0.84 (m, 1H) 1.07, (m, 1H) 1.76-1.86 (m, 2H) 2.72 (m, 2H);3.14 (s, 2H); 3.49 (dd, 1H); 3.87 (dd, 1H); 3.58 (m, 1H); 4.01 (d, 1H);4.14, (d, 1H); 4.26, (d, 1H); 4.35, (d, 1H); 4.90, (m, 1H); 5.08, (s,2H); 5.97, (d, 1H), 7.08, (d, 2H); 7.17, (t, 1H); 7.20-7.40, (m, 17H).

EXAMPLE 187

Compound 187. 100 mg of the resulting compound 54A was treated with 1 mLof 90% aqueous TFA and allowed to stand for 12 h. The mixture wasconcentrated in vacuo and the residue taken up in 10 mL of dry CH₂Cl₂,treated with 65 mg of N-Cbz-L-isoleucine (0.235 mmol), 50 μL of DIEA(0.27 mmoles), 30 mg of HOBt (0.22 mmoles), and 42 mg of1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.22mmoles). The mixture was stirred for 3 h, then diluted with in CH₂Cl₂and washed sequentially with water, saturated NaHCO₃ solution, andbrine. After drying over MgSO₄ and concentrating in vacuo, the mixturewas purified by chromatography on a silica gel column using 5% CH₃OH inCH₂Cl₂ as eluent to yield the title compound, a portion which waspurified by preparative reversed-phase C₁₈ HPLC using a linear gradientof 35% to 100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 36.0 mg99.0% pure compound. TLC: Rf=0.25, 5% CH₃OH in CH₂Cl₂. HPLC: Rt=16.45min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 188

Compound 188. A solution of 51 mg of the resulting compound of Example187A in 15 mL of methanol was hydrogenated under a slight positivepressure of hydrogen in the presence of 10 mg of 10% Pd(OH)₂ for 14 h.After filtering and concentrating in vacuo, the crude mixture was takenup into 10 mL CH₂Cl₂ and treated with 0.203 mL of DIEA and 19.0 mg of2-quinoxaloyl chloride. The mixture was stirred for 6 h, then dilutedwith CH₂Cl₂ and washed with water. After drying over MgSO₄ andconcentrating in vacuo, a portion of the mixture was purified bypreparative reversed-phase C₁₈ HPLC using a linear gradient of 35% to100% CH₃CN/H₂O with 0.1% TFA for elution to obtain 2.1 mg of the titlecompound. TLC: Rf=0.25, 6% CH₃CN/H₂O with 0.1% TFA for elution to obtain2.1 mg of the title compound. TLC: Rf=0.25, 6% CH₃OH in CH₂Cl₂. HPLC:Rt=16.21 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 189

A. Compound XXII (D′=isobutyl, A=H, E=4-acetamidophenyl,trifluoroacetate salt). To a solution of 89.3 mg. (0.167 mmol) of theresultant compound of Example 39B in CH₂Cl₂ (1 mL) at 0° to 5° C. wasadded trifluoromethanesulfonic acid (1 mL). After stirring for 0.5 h theresultant mixture was concentrated in vacuo and the resulting yellow gumused without subsequent purification.

B. Compound 189. A solution of the resultant compound of Example 189A(0.167 mmol) in CH₂Cl₂ was treated sequentially, at ambient temperatureunder an atmosphere of nitrogen, with 44.2 mg (0.217 mmol) ofN-Boc-α-aminoisobutyric acid, 0.044 mL (0.251 mmol)diisopropylethylamine, 27.1 mg (0.201 mmol) of 1-hydroxybenzotriazolehydrate, 38.5 mg (0.201 mmol)1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride. The mixturewas stirred for 16 h and then concentrated in vacuo. The residue wastaken up in ethyl acetate and washed with water, 0.5 N hydrochloricacid, washed with sodium bicarbonate, saturated brine, dried overmagnesium sulfate, filtered and concentrated in vacuo. The residue waspurified by low pressure silica gel column chromatography using a 10% to35% gradient of ethyl acetate/CH₂Cl₂ eluent to yield 69.3 mg of thetitle product as a white solid. TLC: Rf=0.46, 60% ethyl acetate/CH₂Cl₂,HPLC: Rt=15.0 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 190

A. Compound XXXI (A=H, R³=methyl, R^(3′)=methyl, D′=isobutyl,E=4-acetamidophenyl, hydrochloride salt). To a solution of 60.1 mg ofthe resultant compound of Example 189B in CH₂Cl₂ (1 mL) at 0° to 5° C.was added trifluoromethanesulfonic acid (1 mL). After stirring for 0.75h, the resultant mixture was concentrated in vacuo and the resultingwhite solid used directly for subsequent reaction.

B. Compound 190. To a solution of 37 mg (0.059 mmol) of the resultantcompound of Example 190A in CH₂Cl₂ (3 mL) was added sequentially, atambient temperature under an atmosphere of nitrogen, with 15.4 mg (0.089mmol) of 1-hydroxybenzotriazole hydrate, and 17.8 mg (0.089 mmol) EDC.The mixture was stirred for 16 h and then concentrated in vacuo. Theresidue was taken up in EtOAc and washed with saturated brine, driedover magnesium sulfate, filtered and concentrated in vacuo. The residuewas purified by thin layer silica gel column chromatography using 50% ofEtOAc in CH₂Cl₂ as eluent to yield 32.5 mg of the title product. TLC:Rf=0.35, 50% EtOAc/CH₂Cl₂, HPLC: Rt=15.65 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 191

A. (2S,3RS)-S-Amino-1-chloro-2-hydroxy-4-phenylbutane). A solution of2.24 g (6.71 mmol) of(1S,2RS)-N-(1-benzyl-3-chloro-2-hydroxypropyl)-benzyloxycarbonylamine in5 mL of methanol was added, at ambient temperature under a nitrogenatmosphere, to a slurry of 0.22 g (10% by weight) of 10% palladium oncarbon in 60 mL methanol and hydrogenerated for 24 h, under a slightpositive pressure of hydrogen. The mixture was filtered and concentratedin vacuo to yield 1.34 g of the mixed diastereomeric products. TLC:Rf=0.33, 10% CH₃OH/CH₂Cl₂.

B.(2S)-2-Benzyloxycarbonylamino-N¹-((1S,2RS)-1-benzyl-3-chloro-2-hydroxypropyl)-N⁴-tritylsuccinamide. A solution of 1.34 g (6.71 mmol) of the resultant compoundsof Example 191A in 60 mL of dichloromethane was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 3.58 g (7.05mmol) of Cbz-N^(δ)-trityl-asparagine, 0.95 g (7.05 mmol) of1-hydroxybenzotriazole hydrate, 1.35 g (7.05 mmol) of EDC. The mixturewas stirred for 24 hours and then concentrated in vacuo. The residue wastaken up in ethyl acetate and washed with water, saturated NaHCO₃,saturated NaCl; dried over MgSO₄; filtered and concentrated in vacuo.The residue was purified by low pressure silica gel columnchromatography using 10% ethyl acetate/dichloromethane as eluent toyield 3.08 g total of the mixed diastereomeric products. TLC: Rf=0.75,0.83, 40% EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

C.(2S)-2-Amino-N¹-((1S,2RS)-1-benzyl-3-chloro-2-hydroxypropyl)-N⁴-tritylsuccinamide. A solution of 2.80 g (4.06 mmol) of the resultant compoundsof Example 191B in 5 mL of methanol was added, at ambient temperatureunder a nitrogen atmosphere, to a slurry of 0.28 g (10% by weight) of10% palladium on carbon in 100 mL methanol and hydrogenated for 24 hunder a slight positive pressure of hydrogen. The mixture was filteredand concentrated in vacuo to yield 2.26 g of the mixed distereomericproducts. TLC: Rf=0.42, 10% CH₃OH/CH₂Cl₂

D.(2S)-2-((1S,2RS)-1-Benzyl-3-chloro-2-hydroxypropyl)-N′-((quinoline-2-carbonyl)-amino)-N⁴-tritylsuccinamide. A solution of 2.26 g (4.06 mmol) of the resultant compoundsof Example 191C in 60 mL of dichloromethane was treated sequentially, atambient temperature under an atmosphere of nitrogen, with 0.74 g (4.27mmol) of quinaldic acid, 0.58 g (4.27 mmol) of 1-hydroxybenzotriazolehydrate, and 0.82 g (4.27 mmol) of EDC. After 24 hours, 30 mL ofdichloromethane was added. The mixture was washed with water, 5% NaHCO₃solution, saturated NaCl, dried over MgSO₄, filtered and concentrated invacuo. The residue was dissolved in 50% ethyl acetate/hexane andfiltered through a plug of silica gel. Removal of the solvents yielded2.30 g of the mixed diastereomeric products. TLC: Rf=0.53, 0.58, 40%EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

E.(2S)-2-((1S,2RS)-1-Benzyl-2-hydroxy-3-iodopropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-tritylsuccinamide. A solution of 1.05 g (1.48 mmol) of the resultant compoundsof Example 191D and 0.36 g (2.37 mmol) of sodium iodide in 15 mL ofmethyl ethyl ketone was heated to reflux for 24 hours. The mixture wascooled to room temperature and then concentrated in vacuo. The residuewas taken up in dichloromethane and washed with water, saturated NaCl,dried over MgSO₄, filtered and concentrated in vacuo to yield 1.3 g ofthe mixed diastereomeric products. TLC: Rf=0.58, 0.65, 40% EtOAc/CH₂Cl₂;(¹H)-NMR (CDCl₃) consistent with structure.

F. (2S)-2-((1S,2 syn,anti)-3-(2-methylpropyl)amino-1-benzyl-2-hydroxypropyl)-N¹-((quinoline-2-carbonyl)-amino)-N⁴-tritylsuccinamide. A solution of 207.6 mg (0.26 mmol) of the resultantcompounds of Example 191E and 0.5 mL (5.17 mmol) of isobutylamine in 9mL of acetonitrile in a sealed tube was heated to reflux for 24 hours.After cooling to room temperature, the mixture was concentrated invacuo. The residue was taken up in dichloromethane and washed withwater, saturated NaCl, dried over MgSO₄, filtered and concentrated invacuo to yield 209.2 mg of the mixed diastereomeric products. TLC:Rf=0.11, 10% CH₃OH/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

G. Compound XIV ((syn, anti)-OH, P=quinoline-2-carbonyl, D′=isobutyl). Asolution of 192.9 mg (0.26 mmol) of the resultant compounds of Example191F and 0.07 mL (0.388 mmol) of diisopropylethylamine in 5 mL ofdichloromethane was treated with 112.9 mg (0.517 mmol) ofdi-tert-butyldicarbonate. After 24 hours, the mixture was diluted withdichloromethane. The mixture washed with water, 5% NaHCO₃, 0.5 N HCl,saturated NaCl, dried over MgSO₄, filtered and concentrated in vacuo.The residue was purified by low pressure silica gel columnchromatography using 40% ethyl acetate/dichloromethane as eluent toyield 147.3 mg of the mixed diastereomeric products. TLC: Rf=0.60, 0.67,40% EtOAc/CH₂Cl₂; (¹H)-NMR (CDCl₃) consistent with structure.

H. Compounds 191. A solution of 147.3 mg (0.174 mmol) of the resultantcompounds of Example 191G in 2 mL of dichloromethane was treated with 2mL of trifluoroacetic acid. After 4 hours, the mixture was concentratedin vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solution of the resultantcompound in 2 mL of dichloromethane was sequentially added 0.5 mL ofsaturated NaHCO₃, small amount of solid NaHCO₃ and 67 mg (0.226 mmol) ofa mixture of 4-acetamido-3-fluorobenzenesulphonyl chloride and3-acetamido-4-fluorobenzenesulphonyl chloride. After 3 hours, themixture was diluted with dichloromethane. The two layers were separatedand the aqueous layer was extracted once with dichloromethane. Thecombined organic layer was washed with saturated NaCl then dried overMgSO₄, filtered and concentrated in vacuo. The residue was purified bylow pressure silica gel column chromatography using 2%methanol/dichloromethane was eluent to yield 64 mg of the mixeddiastereomers and regioisomers which were further purified withpreparative HPLC to yield 18.9 mg of the mixed regioisomers comprisingcompounds 191 as a white solid. TLC: Rf=0.14, 5% CH₃OH/CH₂Cl₂; HPLC,Rt=13.36 min; (¹H)-NMR (CDCl₃) consistent with structure.

EXAMPLE 193

Compound 193. A solution of 81.2 mg (0.096 mmol) of the resultant lowerRf diastereomer of Example 9/192A in 3 mL of dichloromethane was treatedwith 3 mL of trifluoroacetic acid. After 4 hours, the mixture wasconcentrated in vacuo. TLC: Rf=0.11, 10% CH₃OH/CH₂Cl₂. To a solution of20.6 mg (0.0431 mmol) of the resultant residue in 1 mL ofdichloromethane was sequentially added 0.3 mL of saturated NaHCO₃, smallamount of solid NaHCO₃ and 12.4 mg (0.053 mmol) of4-acetamidobenzenesulphonyl chloride. After 3 hours, the mixture wasdiluted with dichloromethane. The two layers were separated and theaqueous layer was extracted once with dichloromethane. The combinedorganic layer was washed with brine then dried over MgSO₄, filtered andconcentrated in vacuo. The residue was purified by preparative HPLC toyield 8.3 mg of the title compound as a white solid; TLC: Rf=0.10, 5%CH₃OH/CH₂Cl₂; HPLC, Rt=12.7 min; (¹H)-NMR (CDCl₃) consistent withstructure.

EXAMPLE 194

Compound 194. To a solution of 13.0 mg (0.026 mmol) of thetrifluoroacetic acid deprotection product described in Example 193 in 1mL of dichloromethane was sequentially added 0.3 mL of saturated NaHCO₃,small amount of solid NaHCO₃ and 8.4 mg (0.033 mmol) of5-(isoxazol-3-yl)thiophene-2-sulphonyl chloride. After 3 hours, themixture was diluted with dichloromethane. The two layers were separatedand the aqueous layer was extracted once with dichloromethane. Thecombined organic layer was washed with brine then dried over MgSO₄,filtered and concentrated in vacuo. The residue was purified bypreparative HPLC to yield 5.1 mg of the title product as a white solid;TLC: Rf=0.27, 5% CH₃OH/CH₂Cl₂; HPLC, Rt=14.4 min; (¹H)-NMR (CDCl₃)consistent with structure.

EXAMPLE 195

A. Compound XXII (A=(S)-3-tetrahydrofuryl, D′=cyclopentylmethyl,A′=tert-butoxycarbonyl). To a solution of 264 mg of the resultantcompound of Example 140D in 10 mL of CH₂Cl₂ was added 0.14 mL ofdisopropylethylamine and 175 mg of di-tert butylpyrocarbonate. Afterstirring for 4 hours, the mixture was diluted with 50 mL of CH₂Cl₂,washed with 0.5N of HCl and brine, dried over magnesium sulfate,filtered and concentrated in vacuo to yield 364 mg of the title compoundas a white solid which was used without subsequent purification. TLC:Rf=0.58, 40% EtOAc/CH₂Cl₂.

B. A solution of 334 mg of the resultant compound of Example 195A in 5mL of ethanol was hydrogenated under 30 psi of hydrogen in the presenceof 80 mg of platinum (IV) oxide for 24 hours. The mixture was filteredand concentrated. The residue was purified by a low pressure silica gelcolumn chromatography using 20% EtOAc in CH₂Cl₂ eluent to yield 268 mgof the title compound. TLC: Rf=0.55, 40% EtOAc/CH₂Cl₂. (¹H)-NMR (CDCl₃)consistent with structure.

C. A solution of 268 mg of the resultant compound of Example 195B in 10mL of EtOAc was treated with anhydrous HCl gas for 5 min. The reactionmixture was sparged with nitrogen then concentrated in vacuo and theresulting white solid used without subsequent purification forsubsequent reaction.

D. Compound 195. To a solution of 233 mg of the crude resultant compoundof Example 195C in 10 mL of CH₂Cl₂ was added 2 mL of saturated aqueoussodium bicarbonate and 149 mg of 4-methyloxybenzene sulfonyl chloride.After 3 hours, the resulting mixture was diluted with CH₂Cl₂, washedwith sodium bicarbonate, brine, dried over magnesium sulfate, filteredand concentrated in vacuo. The residue was purified by low pressuresilica gel column chromatography using 0% to 20% EtOAc/CH₂Cl₂ to yield225 mg of the title compound as a white solid. TLC: Rf=0.40, 20%EtOAc/CH₂Cl₂; HPLC: Rt=15.65 min.: (¹H)NMR (CDCl₃) consistent withstructure.

EXAMPLE 196

A.(1S,2S)-N-(1-Isobutyl-3-chloro-2-hydroxypropyl)benzyloxycarbonylamine.To a solution of N-Cbz-leucine chloromethyl ketone (2.0 g) in 20 mL ofmethanol was added, at 0° C., 1.0 g of sodium borohydride and themixture was stirred at ambient temperature for 24 h. The solution wasconcentrated under reduced pressure and the residue partitioned between20 mL of saturated aqueous NH₄Cl and 500 ml of diethyl ether. Theorganic fraction was separated, dried over MgSO₄ and concentrated invacuo and the residue purified by silica gel chromatography to yield 1.8g of white solid.

B. (1S)-1-1(S)(Carbobenzyloxy)amino-2-isobutyl-oxirane. To a solution ofthe resultant compound of Example 196A (300 mg) in absolute ethanol wasadded 67 mg of powdered KOH. The mixture was stirred for 3 h at ambienttemperature, filtered through diatomaceous earth, and concentrated invacuo. The residue was dissolved in diethyl ether, dried over MgSO₄, andconcentrated to yield 230 mg of colorless oil, which was used directlyfor subsequent reaction.

C.(2R,3S)-N³-Carbobenzyloxy-N¹-isobutyl-1,3-diamino-2-hydroxy-5-methylhexane.A 230 mg portion of the resultant compound of example 196B was suspendedin 5 mL of isobutylamine and the mixture stirred overnight at ambienttemperature. The mixture was concentrated in vacuo to yield the titleproduct as 179 mg of a white solid, which was used directly forsubsequent reaction.

D. Compound I (A=tert-butoxycarbonyl, x=0, D=isobutyl,E=4-methoxyphenyl, (s)-hydroxy). Following the procedure described inExample 81, a solution of the resultant compound of example 196C (170mg) in CH₂Cl₂ was reacted with 4-methoxybenzenxulfoyl chloride (150 mg)in the presence of aqueous NaHCO₃. Workup and silica gel chromatographyyielded 90 mg of product as a white solid.

E. Compound I (A=H, x=0, D=isobutyl, E=4-methoxyphenyl, (S)-hydroxy). Asolution of the resultant compound of Example 196D (90 mg) in ethanolwas treated with 50 mg of 10% palladium on carbon and the mixturestirred under an atmosphere of hydrogen. After completion of reaction,the mixture was filtered and concentrated in vacuo to yield 60 mg of thetitle compound which was used directly for subsequent reaction.

F. Compound 196. Reaction of the resultant compound of Example 196E (60mg) in CH₂Cl₂ was reacted with the resultant product of example 48A (150mg) as described earlier yielded, following aqueous workup, drying overMgSO₄, filtering, and concentration in vacuo, a residue which waspurified by silica gel chromatography using methanol/CH₂CL₂ as eluant toyield the title product as 40 mg of white solid. [¹h]-NMR (CDCl₃)consistent with structure.

EXAMPLE 197

We measured the inhibition constants of the compounds listed in TableVII against HIV-1 protease using the above-cited method of Pennington etal.

We also measured the anti-viral potency of the compounds in CCRM-CEMcells by the above-cited method of Meek et al. In the Tables below,K_(i) and IC₉₀ values are expressed in nM.

In Table VIII, the following classifications have been employed:

-   -   A: inhibits HIV replication at concentration of 100 nM or less.    -   B: inhibits HIV replication at concentration of between 101 and        1,000 nM.    -   C: inhibits HIV replication at a concentration of between 1,001        and 10,000 nM.    -   D: inhibits HIV replication at a concentration of between 10,001        and 40,000 nM.    -   ND: not tested.

TABLE VII Compound K_(i)value 1 4.0 2 2.0 3 32 4 19 5 2.0 6 3.0 7 8.0 8850 9 4.0 10 4.0 11 34 12 0.1 13 0.2 14 0.1 15 <0.1 16 <0.1 17 <0.1 18<0.1 19 <0.1 20 0.1 21 0.7 22 1.0 23 1.5 24 32,500 25 3,000 26 0.1 278.0 28 17 29 17 30 61 31 ND 32 2.5 33 80 34 17 35 4.0 36 19 37 0.1 381.5 39 17 40 1,100 41 220 42 46 43 4,200 44 5.0 45 6.0 46 154 47 4.0 481.4 49 9.0 50 11 51 ND 52 0.4 53 27 54 22 55 430 56 60 57 200 58 34 59206 60 4.0 61 4.0 62 72 63 7.0 64 3.0 65 0.7 66 0.4 67 7,400 68 120 6942 70 25 71 470 72 4000 73 140 74 11 75 290 76 ND 77 ND 78 ND 79 ND 80ND 81 2.3 82 1.5 83 ND 84 1.4 85 4.0 86 5.0 87 10 88 1.4 89 2.0 90 93 912.5 92 20 93 0.8 94 1.7 95 1.3 96 8.0 97 2.5 98 0.5 99 0.24 100 0.16 101250 102 33 103 4.5 104 5.5 105 7.5 106 1.4 107 1.4 108 2.0 109 6.0 11028 111 0.3 112 4.0 113 3.0 114 0.35 115 0.5 116 <0.1 117 0.26 118 <0.1119 1.8 120 11 121 2.0 122 1.2 123 10 124 1.1 125 0.3 126 310 127 650128 >5000 129 19 130 14 131 60 132 6.0 133 24 134 8.4 135 2.7 136 18 13726 138 1.4 139 1.2 140 <0.1 141 0.1 142 <0.1 143 <0.1 144 8.0 145 1.4146 2.0 147 1.6 148 0.2 149 1.7 150 6.0 151 0.8 152 2.5 153 0.2 154 0.5155 1.7 156 2.8 157 0.7 158 <0.1 159 0.2 160 1.0 161 20 162 0.5 163 0.5164 130 165 0.4 166 <0.1 167 0.45 168 0.6 169 <0.1 170 0.2 171 0.2 17221 173 0.6 174 10 175 0.1 176 <0.1 177 <0.1 178 0.1 179 0.4 180 <0.1 1810.3 182 0.2 183 0.1 184 5.0 185 3.5 186 140 187 0.3 188 11.5 189 5,500190 ND 191 33 192 67 193 400 194 350 195 0.2 196 ND

TABLE VIII Compound IC₉₀ Range 1 C 2 B 3 C 4 C 5 B 6 B 7 D 8 ND 9 B 10 B11 ND 12 A 13 A 14 A 15 A 16 B 17 B 18 B 19 B 20 A 21 A 22 B 23 B 24 ND25 ND 26 B 27 C 28 ND 29 C 30 ND 31 ND 32 C 33 ND 34 ND 35 B 36 ND 37 B38 C 39 C 40 ND 41 ND 42 ND 43 ND 44 B 45 C 46 ND 47 C 48 B 49 C 50 C 51C 52 B 53 ND 54 C 55 ND 56 ND 57 ND 58 ND 59 ND 60 C 61 C 62 ND 63 C 64C 65 C 66 B 67 ND 68 ND 69 ND 70 ND 71 ND 72 ND 73 ND 74 ND 75 ND 76 ND77 ND 78 ND 79 ND 80 ND 81 C 82 C 83 ND 84 C 85 C 86 B 87 C 88 B 89 C 90ND 91 B 92 ND 93 B 94 B 95 C 96 ND 97 B 98 B 99 B 100 A 101 ND 102 ND103 C 104 C 105 ND 106 C 107 C 108 C 109 B 110 ND 111 C 112 B 113 B 114B 115 B 116 A 117 C 118 B 119 C 120 ND 121 C 122 C 123 ND 124 D 125 B126 ND 127 ND 128 ND 129 ND 130 ND 131 ND 132 ND 133 ND 134 ND 135 C 136ND 137 ND 138 B 139 B 140 A 141 B 142 A 143 A 144 B 145 B 146 B 147 B148 A 149 B 150 B 151 C 152 ND 153 ND 154 ND 155 B 156 B 157 B 158 A 159B 160 A 161 ND 162 C 163 B 164 ND 165 B 166 A 167 B 168 A 169 A 170 B171 A 172 ND 173 A 174 ND 175 A 176 ND 177 ND 178 ND 179 ND 180 ND 181ND 182 B 183 B 184 ND 185 ND 186 ND 187 B 188 C 189 ND 190 ND 191 C 192C 193 ND 194 ND 195 A 196 ND

As demonstrated in Tables VII and VIII, all of the compounds testeddisplayed inhibitory and anti-viral activity. Moreover, several of thesecompounds exhibited activity levels far greater than those of known HIVprotease inhibitors.

While we have described a number of embodiments of this invention, it isapparent that our basic constructions may be altered to provide otherembodiments which utilize the products and processes of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims, rather than by the specificembodiments which have been presented by way of example.

1. A compound of formula I:

wherein: A is selected from the group consisting of H; Het; —R¹-Het;—R′—C₁-C₆ alkyl, which may be optionally substituted with one or moregroups selected from the group consisting of hydroxy, C₁-C₄ alkoxy, Het,—O-Het, —NR²—CO—N(R²)(R²) and —CO—N(R²)(R²); and —R¹—C₂-C₆ alkenyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of hydroxy, C₁-C₄ alkoxy, Het, —O-Het,—NR²—CO—N(R²)(R²) and —CO—N(R²)(R²); each R¹ is independently selectedfrom the group consisting of —C(O)—, —S(O)₂—, —C(O)—C(O)—, —O—C(O)—,—O—S(O)₂, —NR²—S(O)₂—, —NR²—C(O)— and —NR²—C(O)—C(O)—; each Het isindependently selected from the group consisting of C₃-C₇ cycloalkyl;C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7 membered saturated orunsaturated heterocycle, containing one or more heteroatoms selectedfrom N,N(R²), O, S and S(O)_(n), wherein said heterocycle may optionallybe benzofused; and wherein any member of said Het may be optionallysubstituted with one or more substituents selected from the groupconsisting of oxo, —OR², —R², —N(R²)(R²), —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)(R²), —S(O)₂—N(R²)(R²), —N(R²)—C(O)—R₂, —C(O)—R²,—S(O)_(n)—R², —OCF₃, —S(O)_(n)—Ar, methylenedioxy, —N(R²)—S(O)₂(R²),halo, —CF₃, —NO₂, Ar and —O—Ar; each R² is independently selected fromthe group consisting of H and C₁-C₃ alkyl optionally substituted withAr; B, when present, is —N(R²)—C(R³)(R³)—C(O)—; x is 0 or 1; each R³ isindependently selected from the group consisting of H, Het, C₁-C₆ alkyl,C₂-C₆ alkenyl, C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl, wherein anymember of said R³, except H, may be optionally substituted with one ormore substituents selected from the group consisting of —OR²,—C(O)—NH—R², —S(O)_(n)—N(R²)(R²), Het, —CN, —SR², —CO₂R², NR²—C(O)—R²;each n is independently 1 or 2; D and D′ are independently selected fromthe group consisting of Ar; C₁-C₄ alkyl, which may be optionallysubstituted with one or more groups selected from C₃-C₆ cycloalkyl,—OR₂, —R³, —O—Ar and Ar; C₂-C₄ alkenyl, which may be optionallysubstituted with one or more groups selected from the group consistingof C₃-C₆ cycloalkyl, —OR², —R³, —O—Ar and Ar; C₃-C₆ cycloalkyl, whichmay be optionally substituted with or fused with Ar; and C₅-C₆cycloalkenyl, which may be optionally substituted with or fused with Ar;each Ar is independently selected from the group consisting of phenyl;3-6 membered carbocyclic ring and 5-6 membered heterocyclic ringcontaining one or more heteroatoms selected from O, N, S, S(O)_(n) andN(R²), wherein said carbocyclic or heterocyclic ring may be saturated orunsaturated and optionally substituted with one or more groups selectedfrom the group consisting of oxo, —OR², —R², —N(R²)(R²), —N(R²)—C(O)—R²,—R²—OH, —CN, —CO₂R², —C(O)—N(R²)(R²), halo and —CF₃; E is selected fromthe group consisting of Het; O-Het; Het-Het; —O—R³; —NR²R³; C₁-C₆ alkyl,which may be optionally substituted with one or more groups selectedfrom the group consisting of R⁴ and Het; C₂-C₆ alkenyl, which may beoptionally substituted with one or more groups selected from the groupconsisting of R⁴ and Het; C₃-C₆ saturated carbocycle, which mayoptionally be substituted with one or more groups selected from thegroup consisting of R⁴ and Het; and C₅-C₆ unsaturated carbocycle, whichmay optionally be substituted with one or more groups selected from thegroup consisting of R⁴ and Het; and each R⁴ is independently selectedfrom the group consisting of −OR², —C(O)—NHR², —S(O)₂—NHR², halo,—NR²—C(O)—R² and —CN.
 2. The compound according to claim 1, wherein saidcompound has the structure of formula XXII:

and A, D′ and E are defined as in claim
 1. 3. The compound according toclaim 1, wherein said compound has the structure of formula XXIII:

and x, Het, R³, D′ and E are defined as in claim
 1. 4. The compoundaccording to claim 1, wherein said compound has the structure of formulaXXXI:

and A, R³, D′ and E are defined as in claim
 1. 5. A compound accordingto claim 1, wherein: A is selected from the group consisting of H;—R¹-Het; —R¹-C₁-C₆ alkyl, which may be optionally substituted with oneor more groups selected from the group consisting of hydroxy, C₁-C₄alkoxy, Het and —O-Het; and —R¹—C₂-C₆ alkenyl, which may be optionallysubstituted with one or more groups selected from hydroxy, C₁-C₄ alkoxy,Het and —O-Het; each R¹ is independently selected from the groupconsisting of —C(O)—, —S(O)₂—, —C(O)—C(O)—, —O—CO—, —O—S(O)₂— and—NR²—S(O)₂—; each Het is independently selected from the groupconsisting of C₃-C₇ cycloalkyl; C₅-C₇ cycloalkenyl; C₆-C₁₀ aryl; and 5-7membered saturated or unsaturated heterocycle, containing one or moreheteroatoms selected from N, O and S, which may optionally bebenzofused; wherein any member of said Het may be optionally substitutedwith one or more substituents selected from the group consisting of oxo,—OR², —R², —N(R²)₂, —R²—OH, —CN, —CO₂R², —C(O)—N(R²)₂ and —S(O)₂—N(R²)₂;each R² is independently selected from the group consisting of H andC₁-C₃ alkyl; B, when present, is —NH—CH(R³)—C(O)—; x is 0 or 1; R³ isselected from the group consisting of Het, C₁-C₆ alkyl, C₂-C₆ alkenyl,C₃-C₆ cycloalkyl and C₅-C₆ cycloalkenyl, wherein any member of said R³may be optionally substituted with one or more substituents selectedfrom the group consisting of −OR², —C(O)—NH—R², —S(O)_(n)—N(R²)₂, Hetand —CN; n is 1 or 2; D and D′ are independently selected from the groupconsisting of Ar; C₁-C₄ alkyl, which may be optionally substituted withC₃-C₆ cycloalkyl or Ar; C₂-C₄ alkenyl, which may be optionallysubstituted with C₃-C₆ cycloalkyl or Ar; C₃-C₆ cycloalkyl, which may beoptionally substituted or fused with Ar; and C₅-C₆ cycloalkenyl, whichmay be optionally substituted or fused with Ar; with the proviso thatwhen D is attached to N, D may not be methyl or C₂ alkenyl; Ar isselected from the group consisting of phenyl; 3-6 membered carbocyclicring and 5-6 membered heterocyclic ring containing one or moreheteroatoms selected from O, N and S, wherein said carbocyclic orheterocyclic ring may be saturated or unsaturated and optionallysubstituted with one or more groups selected from the group consistingof oxo, —OR², —R², —N(R²)₂, —N(R²)—C(O)R², —R²—OH, —CN, —CO₂R²,—C(O)—N(R²)₂, halo and —CF₃; E is selected from the group consisting ofHet; —O—R³; —NR²R⁵; C₁-C₆ alkyl, which may be optionally substitutedwith one or more R⁴ or Het; C₂-C₆ alkenyl, which may be optionallysubstituted with one or more R⁴ or Het; C₃-C₆ saturated carbocycle,which may optionally be substituted with one or more R⁴ or Het; andC₅-C₆ unsaturated carbocycle, which may optionally be substituted withone or more R⁴ or Het; each R⁴ is independently selected from the groupconsisting of —OR², —C(O)—NHR², —S(O)₂—NHR², halo and —CN; and each R⁵is independently selected from the group consisting of H and R³, withthe proviso that at least one R⁵ is not H.
 6. The compound according toclaim 2, wherein: A is R¹-Het; and D′ is selected from the groupconsisting of C₁-C₃ alkyl and C₃ alkenyl, wherein said alkyl or alkenylmay optionally be substituted with one or more groups selected from thegroup consisting of C₃-C₆ cycloalkyl, —OR², —O—Ar and Ar.
 7. Thecompound according to claim 3, wherein: R³ is selected from the groupconsisting of C₁-C₆ alkyl, C₂-C₆ alkenyl, C₅-C₆ cycloalkyl, C₅-C₆cycloalkenyl and a 5-6 membered saturated or unsaturated heterocycle,wherein any member of said R³ may optionally be substituted with one ormore substituents selected from the group consisting of —OR²,—C(O)—NH—R², —S(O)_(n)N(R²)(R²)₂, Het, —CN, —SR², —C(O)₂R², NR²—C(O)—R²;and D′ is selected from the group consisting of C₁-C₃ alkyl and C₃alkenyl, wherein said alkyl or alkenyl may optionally be substitutedwith one or more groups selected from the group consisting of C₃-C₆cycloalkyl, —OR², —O—Ar and Ar.
 8. The compound according to claim 4,wherein: A is R′-Het; each R³ is independently C₁-C₆ alkyl, which may beoptionally substituted with a substituent selected from the groupconsisting of —OR², —C(O)—NH—R², —S(O)_(n)N(R²)₂, Het, —CN, —SR²,—CO₂R², —NR²—C(O)—R²; and D′ is C₁-C₄ alkyl, which may be optionallysubstituted with a group selected from the group consisting of C₃-C₆cycloalkyl, —OR², —O—Ar and Ar; and E is selected from the groupconsisting of Het, Het-Het and —NR²R³. 9-10. (canceled)
 11. The compoundaccording to claim 1, wherein said compound has a molecular weight lessthan or equal to about 700g/mol.
 12. A compound according to claim 11,wherein said compound has a molecular weight less than or equal to about600g/mol.
 13. A compound having the following structural andphysicochemical characteristics: (1) a first and a second hydrogen bondacceptor moiety, at least one of which is more highly polarizable than acarbonyl, said moieties being the same or different, and being capableof hydrogen bonding with the hydrogen atoms of the flap water moleculeof an HIV aspartyl protease when the compound is bound thereto; (2)substantially hydrophobic moieties which associate with the P_(1i) andP₁′ binding pockets of said HIV aspartyl protease when the compound isbound thereto; (3) a third hydrogen bonding moiety, which may be eithera hydrogen bond donor or acceptor, capable of simultaneously hydrogenbonding to Asp25 and Asp25′ of said HIV aspartyl protease when thecompound is bound thereto; (4) an additional occupied volume of space ofat least 100 Å³ when the compound is bound to the active site of saidHIV aspartyl protease, said space overlapping with the volume of spacethat would be filled by a native substrate of said HIV aspartyl proteaseor a nonhyrolyzable isostere thereof; (5) a deformation energy ofbinding of the compound to said HIV aspartyl protease of not greaterthan 10 kcal/mole; and (6) a neutral or favorable enthalpic contributionfrom the sum of all electrostatic interactions between the compound andthe protease when the compound is bound to said HIV aspartyl protease.14-15. (canceled)
 16. A pharmaceutical composition effective againstviral infection comprising a pharmaceutically effective amount of acompound according to claim 1 and a pharmaceutically acceptable carrier,adjuvant or vehicle.
 17. The pharmaceutical composition according toclaim 16, further comprising an additional anti-viral agent. 18-21.(canceled)
 22. A method for preventing HIV infection in a mammalcomprising the step of administering to said mammal a pharmaceuticallyeffective amount of a pharmaceutical composition according to claim 16.23. A method for treating HIV infection in a mammal comprising the stepof administering to said mammal a pharmaceutically effective amount of apharmaceutical composition according to claim
 16. 24. The methodaccording to claim 22, wherein said step of administering comprises oraladministration or administration by injection.
 25. A method foridentification, design, or prediction of an HIV protease inhibitorcomprising the steps of: (a) selecting a candidate compound of definedchemical structure containing a first and a second hydrogen bondacceptor moiety, at least one of which is more highly polarizable than acarbonyl, said moieties being the same or different; a third hydrogenbonding moiety, which may be either a hydrogen bond donor or acceptor;and at least two substantially hydrophobic moieties; (b) determining alow-energy conformation for binding of said compound to the active siteof an HIV aspartyl protease; (c) evaluating the capability of said firstand second hydrogen bond acceptor moieties to form hydrogen bonds to theflap water molecule of said HIV aspartyl protease when said compound isbound thereto in said conformation; (d) evaluating the capability ofsaid substantially hydrophobic moieties to associate with the P₁ and P₁′binding pockets of said HIV aspartyl protease when said compound isbound thereto in said conformation; (e) evaluating the capability ofsaid third hydrogen bonding moiety to form hydrogen bonds to Asp25 andAsp25′ of said HIV aspartyl protease when said compound is bound theretoin said conformation; (f) evaluating the overlap of the occupied volumeof said compound when said compound is bound to said HIV aspartylprotease in said conformation and the occupied volume of a nativesubstrate of HIV aspartyl protease or a nonhydrolyzable isosterethereof, when said polypeptide is bound to said HIV aspartyl protease;(g) evaluating the deformation energy of binding of said compound tosaid HIV aspartyl protease; (h) evaluating the enthalpic contribution ofthe sum of all electrostatic interactions between said compound and saidHIV aspartyl protease when said compound is bound thereto in saidconformation; and (i) accepting or rejecting said candidate compound asan HIV protease inhibitor based upon the determinations and evaluationscarried out in steps (b) through (h). 26-27. (canceled)
 28. The methodaccording to claim 23, wherein said step of administering comprises oraladministration or administration by injection.